Page 1 Ver 4.3a Petition This user’s manual is for the software to be run on Olympus FLUOVIEW FV300 Confocal Laser Scanning Biological Microscope. To ensure safety, obtain optimum performance and familiarize yourself fully with this product, we recommend that you study this manual thoroughly before operation.
Page 3 CAUTION 1. Reproduction, copying or duplication of a part or all of this software and manual is prohibited. 2. The information described in this manual may be subject to change without notice. Registered Trademarks Microsoft, Microsoft Windows, Excel for Windows are registered trademarks of Microsoft Corporation. Other brand names and product names are trademarks or registered trademarks of their respective owners.
Page 4 FLUOVIEW MANUAL CONFIGURATION FLUOVIEW MANUAL CONFIGURATION The FLUOVIEW system uses two manuals including this “User’s Manual” and the on- screen manual built into the software (“Online Help”). The User’s Manual is composed of the five following volumes and subject matter: I .
Page 5 FLUOVIEW MANUAL CONFIGURATION Appendix E Converting Analysis Data into a Chart Using EXCEL ......IV. E-1 Appendix F User Registration................IV. F-1 Appendix G USER REGISTRATION OF FV300 ...........IV. G-1 Appendix H Change of Default Folder for [File I/O] Panel........IV. H-1 Appendix I List of Functions in the [Active Overlays] Dialog Box ......IV. I-1 Appendix J Hand Switch and Microscope Frame Function Allocation ....IV J-1 V .
Page 6 NOTATIONS IN THIS MANUAL NOTATIONS IN THIS MANUAL This manual complies with the following notations. Notation of Caution, Notes and Tips Notation Description Caution to prevent injuries to the user or damage to the product (including surrounding objects). Note for the user. NOTE Hint or one-point advice for user reference.
Page 7 NOTATIONS IN THIS MANUAL Notation of key operations Notation Description Enter The name of a key is enclosed inside The positive sign (+) expresses the combination of more than one key operation. For example, refers to pressing the key while holding the key down.
Page 9 On This Volume This volume describes the outline of the FLUOVIEW FV300 system. Please read this volume so that you can understand the system before use.
Page 11: Table Of Contents
CONTENTS 1 SYSTEM OVERVIEW 1-1 Principle.................... 1-1 1-2 Features of FLUOVIEW FV300 ............1-2 1-3 Light Path Diagram ................1-3 1-4 System Configuration..............1-4 1-4-1 System Component Units and Their Functions ..........1-4 1-4-2 Block Diagram .....................1-7 1-5 Software Functional Configuration ..........1-10 1-5-1 Function Panel and Display Panel..............1-10...
Page 13: System Overview
Principle 1 SYSTEM OVERVIEW Olympus FLUOVIEW is a confocal scanning type laser reflected fluorescence microscope with high resolution, high contrast and drastically improved resolution in the light axis direction thanks to the use of confocal optics. It offers researchers features for sectioning, 3D construction and time-series observation as well as a variety of image processing and analysis functions.
Page 14: Features Of Fluoview Fv300
SYSTEM OVERVIEW/ Features of FLUOVIEW FV300 1-2 Features of FLUOVIEW FV300 The detector has 12-bit resolution and is capable of detecting very small changes in fluorescence inside cells. The system has high resolution of a maximum 2048 x 2048 pixels. The output video signal is non-interlaced to provide a clear image without flickering.
Page 15: Light Path Diagram
SYSTEM OVERVIEW/ Light Path Diagram 1-3 Light Path Diagram Optical Laser fiber cable Barrier filters –– Beam expander Mirror Light axis adjustment plate Pupil lens Excitation dichroic mirror Condensing lens Imaging lens Confocal aperture turret Objective Custom dichroic mirror Specimen separating fluorescence emission Optical...
Page 16: System Configuration
SYSTEM OVERVIEW/ System Configuration 1-4 System Configuration 1-4-1 System Component Units and Their Functions Microscope BX61 upright microscope set up for fluorescence observation. Upright stand Z motor Support the scan unit. Image monitor Motor for driving the Video monitor for Protects the focusing adjustment knobs displaying laser scanning...
Page 17 SYSTEM OVERVIEW/ System Configuration Microscope BX51WI stage-fixed upright microscope set up for fluorescence observation. Photo 1-2 Combination with BX51WI III . SYSTEM OVERVIEW Page III . 1 - 5...
Page 18 SYSTEM OVERVIEW/ System Configuration Microscope IX81 inverted microscope set up for fluorescence observation. Photo 1-3 Combination with IX Transmitted light detector (optional) Unit for acquiring transmitted images. III . SYSTEM OVERVIEW III . Page 1 - 6...
Page 19: Block Diagram
SYSTEM OVERVIEW/ System Configuration 1-4-2 Block Diagram Galvano control cable COMPUTER Galvano X/Y cables, 2.9 m each MONITOR Dsub15pin 50pin 50pin 50pin I/O cable, 2.9 m Analog signal cable 2.9 m Dsub15pin FV5-LCU FV5-PSU-2 Dsub15pin Laser Control BNC BNC Unit Laser POWER SUPPLY Warning...
Page 20 SYSTEM OVERVIEW/ System Configuration The block diagram in AOTF (FV5-COMBA) use. Galvano control cable COMPUTER Galvano X/Y cables, 2.9 m each MONITOR Dsub15pin 50pin 50pin 50pin I/O cable, 2.9 m Analog signal cable 2.9 m Dsub15pin FV5-LCU FV5-PSU-2 Dsub15pin Laser Control BNC BNC Unit Laser...
Page 21 SYSTEM OVERVIEW/ System Configuration For connection to (AOTF) FV5-COMBA Input and output terminals on FV5-LCU back (Coaxial connector) Service connector For connection to FV5- PSU (PC-SYSTEM), 50 For connection to FV5-PSU pin. (TD) (Dsub 25 pin, male terminal) For connection to FV5-TD For connection to PC (PC- (Dsub 25 pin, male terminal) SYSTEM), 50 pin.
Page 22: Software Functional Configuration
SYSTEM OVERVIEW/ Software Functional Configuration 1-5 Software Functional Configuration This software uses panel-type windows. Usually, it is required to “select a menu then select the command to be executed” in order to execute a function provided by software. With the panel system, a software function can be executed easily by “selecting the panel page tab of the function to be executed”, just like when using a system notebook or file folder.
Page 23: Panel Structure Of The Software
SYSTEM OVERVIEW/ Software Functional Configuration 1-5-2 Panel Structure of the Software This software cannot show the page tabs of all function panels at a glance, but uses scrolling to display the desired page tab. Please use the following list of the panels as reference in scrolling.
Page 24: Icons Executed By Dragging & Dropping
SYSTEM OVERVIEW/ Software Functional Configuration 1-5-3 Icons Executed by Dragging & Dropping This software selects image files and observation methods (dye name) by means of dragging & dropping. This allows simple selection based on an intuitive operation of “selecting an icon (image file or observation method), dragging it to the desired position and dropping it there”.
Page 25: Identification Of Images Depending On The Observation Methods
SYSTEM OVERVIEW/ Software Functional Configuration 1-5-4 Identification of Images Depending on the Observation Methods In many occasions, FLUOVIEW displays image Image Icon Significance icons to allow identification of the observation XZ observation method used when each image is acquired. (See table on the left.) XZ observation, 2-channel mode When the [File I/O], [Tile], [Process], [Analyze] Xt observation...
Page 27 On This Volume This volume describes the operating procedures of the FLUOVIEW FV300 system. “Getting Started FLUOVIEW” contains information on the basic operation flow until acquisition of XY images. “APPLIED OPERATIONS” provides detailed operating procedures of the system. Please read this volume so that you can understand the system...
Page 29 CONTENTS 1 Getting Started FLUOVIEW 1-1 Basic Operations ................1-1 1-1-1 Scan Unit .....................1-1 1-1-2 Microscope ....................1-8 1-1-3 General Mouse Operation Procedures............1-17 1-1-4 Names of Major Panel and Window Controls and Their Functions.....1-18 1-2 Outline of LSM Observation Procedures........1-20 1-2-1 Turning Power On..................1-22 1-2-2 Starting the Software .................1-26 1-2-3 Focusing on the Specimen.................1-28...
Page 30 CONTENTS 1-2-12-9 Setting a Lower Scan Speed................. 1-51 1-2-12-10 Stopping Repeated Scanning..............1-52 1-2-13 Adjusting the Detection Light Axis............1-52 1-2-14 Acquiring Image ..................1-52 1-2-15 Saving Image ...................1-54 1-2-16 Exiting from the Software .................1-55 1-2-17 Turning Power Off..................1-56 1-3 Online Help..................1-57 1-3-1 Referencing Method...................1-57 1-3-2 Setup of Microscope and Scan Unit ............1-58 1-3-2-1 Configuring the Microscope................
Page 31 CONTENTS 2-2-2-4 XYZ Observation Mode..................2-41 2-2-2-5 XYT Observation Mode..................2-46 2-2-2-6 XYZT Observation Mode .................2-49 2-2-3 Differences in Image Acquisition Method Between Fluorescent and Transmitted Images ...................2-56 2-2-3-1 Monochrome Image ..................2-56 2-2-3-2 Dual-Fluorochrome Image ................2-60 2-2-3-3 Transmitted Image ...................2-63 2-2-4 Image Acquisition by Rotating It (Rotation Scan) ........2-67 2-2-5 Image Acquisition of Only the Rectangular Position (Clip Scan) ....2-68 2-2-6 Image Acquisition by Magnifying the Rectangular Position (Zoom-In Scan)2-70 2-2-7 High-Speed Image Acquisition..............2-73...
Page 32 CONTENTS 2-3-7-2 Reading the Region File................2-142 2-4 Protocol processor..............2-145 2-4-1 Starting the Protocol Processor..............2-146 2-4-2 Editing the Protocol..................2-148 1 Description of Setting Items ..................2-149 2 Command List......................2-155 3 Protocol Repetition Processing................2-159 4 Input supporting function..................2-160 5 Protocol procedure supporting function ..............
Page 33 CONTENTS 2-5-5-2 Decreasing the Number of Divided Images ...........2-200 2-5-6 Switching the Display Method of Multiple Images........2-202 2-5-7 Displaying Multiple Image Slices Together..........2-203 2-5-7-1 Displaying Multiple Images Per Channel ............2-204 2-5-7-2 Displaying Images of Two Channels Together ..........2-205 2-5-7-3 Displaying Time-Lapse Images..............2-206 2-5-7-4 Displaying Multiple Cross-Section Images.............2-207 2-5-7-5 Displaying Same Images in Different Display Methods .........2-207 2-5-7-6 Re-arranging Images Using the Same Display Method.........2-209...
Page 34 CONTENTS Extract image data from an image data set ............... 2-251 2-7 Image Analysis................2-257 2-7-1 Checking the Intensity of a Specific Part ..........2-258 2-7-1-1 Intensity Values on a Line (Line Profile) ............2-258 2-7-1-2 Intensity Values on a Planar Region (Bird’s Eye View) ......... 2-261 2-7-2 Checking the Intensity Distribution of a Specific Part .......2-265 2-7-2-1 Intensity Distribution on a Line (Histogram) ..........
Page 35 CONTENTS 2-12-1 Displaying the Image Intensity ...............2-304 2-12-3 Displaying the X-coordinate/Y-coordinate of the Image ......2-305 2-12-4 Drawing a Figure in Image ..............2-306 2-12-5 Drawing a Scale in Image ..............2-307 2-12-6 Drawing an Arrow in Image..............2-309 2-12-7 Drawing Color Bars in Image ..............2-310 2-12-8 Deleting Comment .................2-311 2-12-9 Moving Comment ...................2-311 2-12-10 Changing the Comment Size ...............2-312...
Page 36 CONTENTS Appendix E Converting Analysis Data into a Chart Using EXCEL Appendix F User Registration Appendix G USER REGISTRATION OF FV300 Appendix G-1 User Registration............G-1 Appendix G-2 Logging into the FV300 ..........G-3 Appendix G-3 Deleting a User.............. G-4 Appendix H Change of Default Folder for [File I/O] Panel Appendix I List of Functions in the [Active Overlays]...
Page 37 CONTENTS Appendix J Hand Switch and Microscope Frame Function Allocation Appendix J-1 Hand Switch Functions..........J-1 Appendix J-2 Microscope Frame Functions........J-2 Appendix J-2-1 BX ....................J-2 Appendix J-2-2 IX....................J-3 Appendix J-2-3 Focus Adjustment Knob ...............J-4 1 BX 2 IX...
Page 39: Getting Started Fluoview
Getting Started FLUOVIEW/ Basic Operations 1 Getting St arted FLUOVIEW 1-1 Basic Operations 1-1-1 Scan Unit (1) CONFOCAL APERTURE turret (2) DETECTION MODE slider (4) Excitation DM selector knob (3) BARRIER FILTERS slider (1) Confocal aperture switching The confocal aperture includes 5 positions, any of which can be selected by rotating the turret.
Page 40 Getting Started FLUOVIEW/ Basic Operations 1: 60 m The confocal aperture sizes recommended for objectives 2: 100 m are 1, 2 or 3 as shown in the following table. 3: 150 m Use these sizes when the observation image is dark and a 4: 200 m brighter image is required.
Page 41 Getting Started FLUOVIEW/ Basic Operations Detection mode setting The detection modes can be switched over by pushing or pulling the DETECTION MODE slider (2). This switch allows you to set the detection mode of the 2 or 1 fluorescence channel by selecting one of the 3 positions.
Page 42 Getting Started FLUOVIEW/ Basic Operations Laser Slider in the pushed-in position Optical fiber cable Barrier filters Beam expander Mirror Light axis adjustment plate Conde nsing lens Excitation dichroic mirror Pupil lens Image-forming CONFOCAL APERTURE turret lens Slider pushed-in harf way to the position Objective Custom dichromatic Specime...
Page 43 Getting Started FLUOVIEW/ Basic Operations Barrier filter setting The barrier filters can be switched over by pushing or pulling the BARRIER FILTERS slider (3). Up to 2 barrier filters can be mounted for each of CH1 and CH2. The filters are engaged in the light path when the slider switch is pushed in and disengaged when it is pulled out.
Page 44 Getting Started FLUOVIEW/ Basic Operations Kr/Ar laser line filter switching (Available only with the Kr/Ar laser combination) The 488 nm and 568 nm excitation wavelengths of the Kr/Ar laser can be selected according to applications. This switch has 5 positions which can be selected with a rotary knob.
Page 45 Getting Started FLUOVIEW/ Basic Operations FITC TRITC (Attenuates the excitation intensity to reduce leakage of the FITC emission CH2) wavelength / nm OPERATION INSTRUCTIONS Page IV . 1 - 7...
Page 46: Microscope
Getting Started FLUOVIEW/ Basic Operations 1-1-2 Microscope The following figure shows the major controls of a microscope. The configuration of the modules including the specimen stage, revolving nosepiece and lighting equipment may differ from those shown below. For detailed operation procedure of the microscope, refer to the instruction manual of your microscope.
Page 47 Getting Started FLUOVIEW/ Basic Operations (1) Light path selector knob Select the light path between direct observation and laser microscopy. Set to the pushed-in position for direct observation. Set to the pulled-out position for laser microscopy. (2) Cube turret Select the fluorescence observation tube by rotating the turret. Engage the desired cube in the light path for direct fluorescence observation.
Page 48 Getting Started FLUOVIEW/ Basic Operations (5) Filters These filters are used to adjust transmitted light. Be sure to disengage any filter from the light path for transmitted observation using laser. Leaving a filter engaged in the light path will degrade the image quality.
Page 49: Combination With Ax
Getting Started FLUOVIEW/ Basic Operations (7) TV adapter light path selector Combination with AX knob Combination with FVX-LT, this knob is not attached. (1) Light path selector knob (2) Universal vertical illuminator mirror cube (3) Analyzer housing AX-AN(optional) AX-URBC (4) Transmitted light DIC slider U-DICR(optional) (6) Universal condenser U-UCDB...
Page 50 Getting Started FLUOVIEW/ Basic Operations Light path selector knob Select the light path between direct observation and laser microscopy. Set to the pushed-in position for direct observation. Set to the pulled-out position for laser microscopy. Universal vertical illuminator mirror cube housing The turret for the cube for reflected light observation can be switched with the hand switch for transmitted/reflected light.
Page 51 Getting Started FLUOVIEW/ Basic Operations With transmitted light differential interference observation NOTE using an immersion objective, set the microscope’s field diaphragm so that it circumscribes the field of view. Otherwise the contrast may degrade. (This applies to both direct observation and laser differential interference observation.) Filters These filters are used to adjust transmitted light.
Page 52 Getting Started FLUOVIEW/ Basic Operations Combination with IX81 FVF (6) Filters (5) Condenser (4) Transmitted light DIC slider U-DICTS (optional) (1) Light path selector button (2) Cube turret (Analyzer IX2-MDICT Optionally installed.) (8) Hand switch U-HSTR2 (U-FH is optionally available.) OPERATION INSTRUCTIONS Page IV .
Page 53 Getting Started FLUOVIEW/ Basic Operations (1) Light path selector button Select the light path between laser microscopic observation and direct observation. When LED is illuminated, the light path is switched for laser microscopic observation. When LED is not illuminated, the light path is switched for direct observation. (2) Fluorescence mirror unit Select the fluorescence observation tube by rotating the turret.
Page 54 Getting Started FLUOVIEW/ Basic Operations (5) Condenser, polarizing plate Used for transmitted lighting. In addition, the rotary turret for the transmitted light DIC slider and the polarizing plate for differential interference observation (analyzer) are also provided. To perform differential interference observation, engage the transmitted light DIC slider (optional) matching the objective in use in the light path.
Page 55: General Mouse Operation Procedures
Getting Started FLUOVIEW/ Basic Operations 1-1-3 General Mouse Opera tion Procedures Use the mouse to select a command, character string or button. Use the left button of the mouse unless otherwise specified. To select or execute something: Clicking To click the mouse, place the mouse pointer on the desired function and press the mouse button once.
Page 56: Names Of Major Panel And Window Controls And Their Functions
Getting Started FLUOVIEW/ Basic Operations 1-1-4 Names of Major Panel and Window Controls and Their Functions The window as shown below is displayed when FLUOVIEW starts up. FLUOVIEW uses panel-type windows. This section describes the names of the major controls displayed in panels and windows by taking the [Acquire] panel and [Microscope Configuration] window as examples.
Page 57 Getting Started FLUOVIEW/ Basic Operations Check box Clicking this box enables or disables the indicated item. The item is enabled when the check box is checked (x). Minimize button< > Title bar Control menu box Click to turn the window Clicking this box displays a control Shows the title of the window.
Page 58: Outline Of Lsm Observation Procedures
Getting started FLUOVIEW/ Outline of LSM Observation Procedures 1-2 Outline of LSM Observation Procedures Fluorescence observation procedure Start the system. Turn the system power ON. Change the ND filter with a filter (Sec 1-2-1) Start the FLUOVIEW software. with higher transmittance (Sec 1-2-10) (Sec 1-2-2...
Page 59 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures Transmitted light observation procedure Start the system Turn the system power ON. (Sec 1-2-1) Start the FLUOVIEW software. Change the with a filter with higher transmittance. (Sec 1-2-10) ( Sec 1 -2-2) Set the observation condition.
Page 60: Turning Power On
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-1 Turning Power On Set the power switch of each unit to ON. When it is necessary only to read data, it is not necessary to turn the laser power supply and reflected light power supply units ON. To stabilize the laser beam output, we recommend leaving the system for warm-up after turning the laser power ON.
Page 61 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures • Turning the laser power ON Ar laser power supply Ar laser, Multiline Ar laser (type 176B and type 300): (type 176B) (1) Set the power switch to ON. (2) Turn the key to the ON position. (The laser fan will start.) After the key is set to ON, it takes tens of seconds before the laser oscillation starts.
Page 62 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures HeCd laser. (1) Turn the rocker switch(1) of laser controller to “I” (ON). (2) Turn the key switch(2) of laser controller to the ON position. (3) Turn the key switch(3) of remote interface module to the ON position.
Page 63 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures Rotary switch Change right side rotary switch only. Do not touch left side rotary switch. If you change left rotary switch, you may not be able to get optimum performance of this system. (In case of LD405) When you use LD405, align arrow mark of right side rotary switch with When you use LD440, align arrow mark of right side rotary switch with...
Page 64: Starting The Software
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-2 Starting the Software NOTE Before starting up this software, wait for more than 2 minutes after turning the microscope and power supply unit ON. Allow the microscope and power supply unit to initialize themselves for about 2 minutes after they are turned on.
Page 65 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures The following window appears when the FLUOVIEW software starts. Fig. 1-3 Window at Start-up OPERATION INSTRUCTIONS Page 1 - 2 7...
Page 66: Focusing On The Specimen
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-3 Focusing on the Specimen 1-2-3-1 Combination with BX Select the light path for 100% eyepiece by pushing in the light path selector knob (1) on the trinocular tube fully to the stop position.
Page 67: Combination With Ix
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures If you want to use a differential interference unit in transmitted light observation, refer to the instruction manual of your microscope. NOTE With transmitted light differential interference observation using an immersion objective, set the microscope’s field diaphragm so that it circumscribes the field of view.
Page 68 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures Focus on the specimen by looking into the eyepiece. Be sure to adjust the diopter of the eyepiece in advance. (Refer to the instruction manual of the IX70/81 microscope.) When the Manual microscope and the Z motor are in use, clear the check mark in the [Locked] check box in the [Z Stage] sub-panel in the [Acquire] panel (see section 2-2-1-3-7 of this volume), then focus on the specimen by operating the fine...
Page 69: Setting The Lsm Light Path
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-4 Setting the LSM Light Path 1-2-4-1 Combination with Upright Microscope (BX) Pull out the light path selector on the trinocular tube fully to the stop position. Pull out the light path selector (7) on the TV adapter fully to the stop position. Push the hand switch button to (8) set to be displayed in the cube display window (2) on the reflected light fluorescence vertical illuminator (when using the...
Page 70 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures (7) Light path selector knob TV,Photo turret Cube /Cube display window (1) Light path selector knob (This figure shows the cube display window.) (3) Analyzer U-MDICT3 (4) Transmitted DIC slider U-DICTS/WI-DICTHRA (when using BX61WI) (optional) condenser Universal...
Page 71: Combination With Erected Microscope(Ax)
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-4-2 Combination with Erected Microscope(AX) Pull out the light path selector (1) on the trinocular eyepiece fully to the stop position. Pull out the light path selector (7) on the TV adapter fully to the stop position. Select the cube in the cube cassette (2) for the vertical illuminator by pressing the required cube conversion button (8) on the U-HSTR hand switch.
Page 72: Combination With Inverted Microscope (Ix81)
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-4-3 Combination with Inverted Microscope (IX81) From the page tabs on the bottom right of the [Acquire] panel, select the [Settings] sub-panel. Fig. 1-5 [Settings] Sub-panel Select the <LSM> button in the [Light Path] group box. The <LSM>...
Page 73 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures (5) Filters (4) Condenser (3) Transmitted DIC slider U-DICTS (optional) (1) Fluorescence mirror unit ((2)Analyzer IX-MDICT Optionally installed.) (6) Hand switch U-HSTR2 (U-FH is optionally available.) OPERATION INSTRUCTIONS Page 1 - 3 5...
Page 74: Combination With Inverted Microscope (Ix70)
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-4-4 Combination with Inverted Microscope (IX70) Turn the light path selector (1) to Set the intermediate magnification knob (7) to 1X. (The 1.5X position cannot be used.) Rotate the cube turret of the reflected light fluorescence unit to When the IX-AN analyzer (3) is in use, disengage it from the light path by setting the switch to the pulled-out position.
Page 75 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures (5) Filters (6) Condenser (4) Transmitted DIC slider U-DICT (optional) (7) Intermediate magnification knob Left side (3) Analyzer IX-AN (1) Light path selector (optional) (2) Cube turret OPERATION INSTRUCTIONS Page 1 - 3 7...
Page 76: Setting The Dyeing Methods
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-5 Setting the Dyeing Methods From the page tabs on the bottom right of the [Acquire] panel, select the [Dyes] sub-panel. 2. Select the specimen dyeing method by dragging [FITC] and [TRITC] in the [Available Dyes] list box in the [Selected Dyes] group box to the field immediately above the list box.
Page 77 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures One Point! The [Assign dyes manually] check box can also be used to set the dyeing method to the desired channel. Check the [Assign dyes manually] check box in the [Dyes] sub-panel. Select the dyeing method in the [Available Dyes] list box and drag it directly to the field of the [Ch] check box.
Page 78: Selecting The Confocal Aperture
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-6 Selecting the CONFOCAL APERTURE Set the CONFOCAL APERTURE knob (1) to select the optimum confocal aperture number for the objective, that is displayed on the control panel. (Refer to section 1-1-1, “Scan Unit” in this volume.) 1-2-7 Selecting the DETECTION MODE Set the DETECTION MODE slider to the optimum position according to the dye used with the observed specimen.
Page 79 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures Be sure to set the switch to the fully pulled-out or pushed-in position. If the optimum setting cannot be identified, see section 1-3-2-3, “Configuring the Filters” in this volume and follow instructions in the [Microscope Configuration] window. The following table shows the possible combinations of the barrier filters and excitation dichroic mirrors.
Page 80 (1), (2), and (3) of the beam splitter are equipped as the factory configuration. Up to two types of barrier filter can be equipped per channel. If the equipment of another filter set for laser configuration is required, please contact your local Olympus representative. (2) DETECTION MODE slider (5) Filter slider...
Page 81: Setting The Excitation Dm
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-9 Setting the Excitation DM (4) Excitation DM selector knob (1) CONFOCAL APERTURE knob (2) DETECTION MODE slider (3) BARRIER FILTERS slider Using the excitation DM selector knob (4), select the excitation DM according to the wavelength of laser in use.
Page 82: Selecting The Laser Line Filters (Combination Using The Kr/Ar Laser)
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-11 Selecting the Laser Line Filters (Combination Using the Kr/Ar Laser) Select the laser line filters using the laser line filter turret. Reference Examples Laser Line Filters FITC Lucifer Yellow, etc. FITC+TRITC FITC+PI, etc.
Page 83: Setting The Channels
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-12-3 Setting the Channels In the Channel 1 group box, check the check box showing the applicable dyeing method to make the image acquisition ready. In the Channel 2 group box, check the check box showing the applicable dyeing method to ready the image acquisition.
Page 84: Setting The Highest Scan Speed
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-12-4 Setting the Highest Scan Speed 1. Set the scan speed to the fastest speed by using the scale in the [Scan Speed] group box in the [Acquire] panel [Scan Speed] group box Set the scan speed by clicking a point on the scale line.
Page 85: Setting The Xy Observation Mode
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-12-5 Setting the XY Observation Mode In the [Mode] group box in the [Settings] sub-panel, select the [Surface] option button. In the [Mode] group box in the [Acquire] panel, select [800 by 600] from the [Size] drop-down list.
Page 86: Setting The Multiple Sections To Be Observed
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-12-7 Setting the Multiple sections to be Observed While acquiring image, move the Z stage to select the multiple sections to be observed. From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Z Stage] sub-panel.
Page 87: Setting The Area To Be Observed
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-12-8 Setting the Area to be Observed When the observation targets are concentrated in a narrow area or when it is necessary to observe the detail of a specific area, the image of a limited area can be acquired selectively.
Page 88: Stopping Repeated Scanning
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-12-10 Stopping Repeated Scanning After the brightness and gain have been adjusted, select the <STOP SCAN> button in the [Acquire] panel to stop scanning temporarily. 1-2-13 Adjusting the Detection Light Axis Adjust the detection light axis for efficient acquisition of the detection light. This adjustment is particularly important after the excitation DM has been switched using the excitation DM selector knob because this operation tends to deviate the light axis.
Page 89 Getting Started FLUOVIEW/ Outline of LSM Observation Procedures Fig. 1-6 Image Acquired in the [Live] Panel OPERATION INSTRUCTIONS Page 1 - 5 1...
Page 90: Saving Image
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-15 Saving Image Display the [File I/O] panel. When saving images acquired with more than one channel, it is possible to select whether images from more than one image are saved simultaneously or only one of the images is saved.
Page 91: Exiting From The Software
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures Enter the file name in the [File name:] text box. Select “FLUOVIEW MultiTif” from [Save as type:] Select the <Save> button. 1-2-16 Exiting from the Software Click the <Exit> button in the toolbar at the bottom of the screen. This will allow you to exit the software.
Page 92: Turning Power Off
Getting Started FLUOVIEW/ Outline of LSM Observation Procedures 1-2-17 Turning Power Off 1. Turn off the power supply to the units (reflected light power supply, power outlet unit). 2. Turn the laser power OFF. • Ar laser, Multiline Ar laser: Turn the key to the OFF position.
Page 93: Online Help
Getting Started FLUOVIEW/ Online Help 1-3 Online Help The FLUOVIEW application comes with the online help facility which allows you to reference the function and operating procedure description while controlling the application. This section describes a simple method of displaying and consulting online help. 1-3-1 Referencing Method The figure below shows the initial display (table of contents) of the FLUOVIEW Online Help window.
Page 94: Setup Of Microscope And Scan Unit
Getting Started FLUOVIEW/ Online Help One Point! Select the <Contents> button to the initial display. Select the <Back> button to return to the previous information page. 1-3-2 Setup of Microscope and Scan Unit The microscope and scan unit can be set up from the FLUOVIEW software, by selecting the observation method and following the displayed guidance information.
Page 95 Getting Started FLUOVIEW/ Online Help When the dyeing method is selected from the [Available Dyes] list box and the <Apply> button is clicked, the dyeing method will be set automatically to the optimum channels. The pinhole diameters are also automatically set to the optimum channels according to the wavelength and the objective selected in the [Acquire] panel.
Page 96 Getting Started FLUOVIEW/ Online Help One Point! The [Assign dyes manually] check box can also be used to set the dyeing method to the desired channel. Check the [Assign dyes manually] check box in the [Dyes] sub-panel. Select the dyeing method in the [Available Dyes] list box and drag it directly to the field of the [Ch] check box.
Page 97: Configuring The Microscope
Getting Started FLUOVIEW/ Online Help 1-3-2-1 Configuring the Microscope Display the [Acquire] panel. [Settings]/[Z Stage]/[Time Series]/[Dyes]/ [Lasers] sub-panels These are used to set the information required image acquisition. OPERATION INSTRUCTIONS Page 1 - 5 9...
Page 98 Getting Started FLUOVIEW/ Online Help From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Settings] sub-panel. <Scope Control> button Displays useful information for the system setup. Fig. 1-9 [Settings] Subpanel Select the <Scope Control> button. The window as shown below will appear. [Filters] box This represents the filters.
Page 99: Configuring The Scan Unit
Getting Started FLUOVIEW/ Online Help Select the <BI> button in the [Observation] group box, and select the microscopy from the option buttons below it. The system setting points to be changed for <BI> button microscope observation will blink in red. Change the system configuration (setup of light path selector, etc.) by following the guidance given by the red blinking light.
Page 100: Configuring The Filters
Getting Started FLUOVIEW/ Online Help 1-3-2-3 Configuring the Filters The barrier filters, excitation filter and beam splitter are set automatically to the light path according to the dyeing method selected for the specimen. Use the following procedure to change these filters. From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Settings] sub-panel.
Page 101 Getting Started FLUOVIEW/ Online Help Click the <SU Control> button at the bottom of the panel. The window as shown below will appear. [Laser Unit] group box Shows the type of laser to be used. (With the laser combiner operation, the laser type is set and displayed automatically.) When the <On>...
Page 102: Configuring The Microscope (Combination With Bx51, Bx61, Ix81)
Getting Started FLUOVIEW/ Online Help 1-3-2-4 Configuring the Microscope (Combination with BX51, BX61, IX81) When the combination with BX51, BX61, or IX81 is in use, the microscope and scan unit (FV500) can be configured on the FLUOVIEW software. Use the following procedure to configure the microscope. From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Settings] sub-panel.
Page 103 Getting Started FLUOVIEW/ Online Help Select the <Scope Control> button on the bottom of the [Settings] sub-panel. The [Microscope Control panel] window as shown below appears. [EPI lamp] Indicates the EPI lamp. [Shutter] group box Clicking inside the box switches the EPI shutter to be closed/opened, [Filter Turret] group Clicking the filter for...
Page 104 Getting Started FLUOVIEW/ Online Help [Lamp] group box Clicking inside the box switches the TD lamp ON/OFF. The TD lamp is set to OFF. The TD lamp is set to ON. [Options] group box Used for optional IX settings. <Link Setting> button Selects the function to change settings corresponding to the change of IX settings.
Page 105 Getting Started FLUOVIEW/ Online Help Clicking the <Link Setting> button displays the [Link Setting] dialog box as shown below. Checking here links the objective in the [Nosepiece] group box with the condenser turret. Checking here escapes the stage or revolving nosepiece when the objective is selected and changed in the [Nosepiece] group Checking here disengages the top lens from the light path when the objective of 4 or lower magnification is selected in...
Page 106 Getting Started FLUOVIEW/ Online Help Open the setting Open the file to which the microscope settings were exported with the <Export Setting> button. The Settings configured by another user or configured for another combination can be applied. Selecting the <Import Setting> button of the [Link Setting] dialog box displays the [Open] dialog box as shown below.
Page 107 Getting Started FLUOVIEW/ Online Help To change the save destination drive or directory, use the [Save in: ] drop-down list. Enter the setting file name into the [File name] text box. Select the <Save> button to close the dialog box. The <Focus Setting>...
Page 108: Parfocality Correction And Jog Sensitivity Adjustment
Getting Started FLUOVIEW/ Online Help 1-3-2-5 Parfocality Correction and Jog Sensitivity Adjustment (When the BX or IX is used) When the system use the BX or IX microscope, the parfocality correction and jog sensitivity can be set per objective. Open the [Microscope Control Panel] window. For the method of displaying this window, see steps 1 and 2 in section 1-3-2-4, “Configuring the Microscope (Combination with BX51, BX61, IX81)”...
Page 109 Getting Started FLUOVIEW/ Online Help The following option button menu is displayed in the [Pfcl Setup Wizard] group box. If you want to begin the parfocality correction value setting with the objective with the maximum magnification, click the [Switch to...] option button then click the <Next>...
Page 110 Getting Started FLUOVIEW/ Online Help The objective set as the specified in the [Objectives] group box is selected, and the [Pfcl Setup Wizard] group box displays the message shown below. Bring the specimen in focus by observing it visually or on the scanned image, and then click the <Next>...
Page 111 Getting Started FLUOVIEW/ Online Help The [Pfcl Setup Wizard:] group box displays the option button menu shown below. If you want to set the parfocality correction values of objectives with other power values, simply click the <Next> button. Click to proceed to the setting of the objective with next higher power to the current objective.
Page 112 Getting Started FLUOVIEW/ Online Help The [Pfcl Setup Wizard] group box displays the option button menu shown below. Adjust the focal position and click the <Next> button. Click to set the current focal position as the parfocality correction value for the current objective and proceed to the setting of the next objective.
Page 113 Getting Started FLUOVIEW/ Online Help Set the parfocality correction values of objectives by repeating steps 6 and 7 for each. The [Pfcl Setup Wizard] group box displays the option button menu shown below. Click the <Pfcl> button to activate the parfocality correction. [Jog Fine] group box Select the jog sensitivity of each...
Page 115: Applied Operations
APPLIED OPERATIONS /General Operation Procedure 2 APPLIED OPERATIONS 2-1 General Operation Procedure This section describes the general image acquisition procedure with the aim to get accustomed with the operation. Begin using the FLUOVIEW system by acquiring an image or opening an image from a file.
Page 116 APPLIED OPERATIONS /General Operation Procedure Turn power ON and start the FLUOVIEW software. (Sections 1-2-1 & 1-2-2) Acquire an image. (According to the selected Open an image in a file. observation mode) (Section 2-3-2) For detailed operation procedures for image acquisition, see section 2-1-1, “Image Acquisition Procedure (Section A)”.
Page 117: Image Acquisition Procedure (Section (A))
APPLIED OPERATIONS /General Operation Procedure 2-1-1 Image Acquisition Procedure (Section (A)) This section describes the procedure for acquiring images. See sections 2-2 and after for the actual operation methods. The detailed operation methods of each item in the procedure are described in the section specified in parentheses (( )). After “Turn power ON and start the FLUOVIEW software”,...
Page 118: Image Acquisition Procedure In An Observation Mode (Section (B))
APPLIED OPERATIONS /General Operation Procedure 2-1-2 Image Acquisition Procedure in an Observation Mode (Section (B)) As an example of “Acquire an image in an observation mode”, this section describes the procedure in the XY observation mode. For the procedures in other observation modes, see section 2-2-2, “Image Acquisition in Other Observation Modes”...
Page 119: Examples Of Operation Procedures
APPLIED OPERATIONS /General Operation Procedure 2-1-3 Examples of Operation Procedures Begin using the FLUOVIEW system by acquiring an image or opening an image in a file. The procedures for the subsequent operations such as image processing are not in question here. For the detailed operation method of each item in the procedure, see the section specified in parentheses (( )).
Page 120 APPLIED OPERATIONS /General Operation Procedure Example 3) To acquire an image and compared it with Example 4) To open an image in a file, improve its a previously acquired image: contrast and create a presentation image by entering comment, etc. Turn power ON and start the Turn power ON and start the FLUOVIEW software.
Page 121: Image Acquisition
APPLIED OPERATIONS /Image Acquisition 2-2 Image Acquisition Confirm the image to be acquired using the microscope, and acquire its image using FLUOVIEW. The image can be saved in a file as required. NOTE Do not move FLOUVIEW FV300 Window while acquiring an image. OPERATION INSTRUCTIONS Page IV .
Page 122: Image Acquisition In Xy Observation Mode
APPLIED OPERATIONS /Image Acquisition 2-2-1 Image Acquisition in X Y Observation Mode This section describes the basic operation procedure from the system configuration to the image acquisition in the XY observation mode and image saving in a file as shown in the following chart.
Page 123: Configuring The Microscope
APPLIED OPERATIONS /Image Acquisition 2-2-1-1 Configuring the Microsc ope Set the light path so that the image can be observed through the microscope. Display the [Acquire] panel. <Focus> button The repetition images <Once> button can be acquired at a high speed. Acquires an image in the currently selected observation mode and display Use this button to find an optimum image.
Page 124 APPLIED OPERATIONS /Image Acquisition Setting the dyeing method From the page tabs on the bottom right of the [Acquire] panel, select the [Dyes] sub-panel. Place the pointer on the icon displayed [Assign dyes manually] check box [Selected Dyes], and the Checking this enables the manual dyeing method is shown in setting.
Page 125 APPLIED OPERATIONS /Image Acquisition When a dyeing method is selected from the [Available Dyes] list box and the <Apply> button is clicked to apply it in the image acquisition channel, the dyeing method emitting a shorter light wavelength than 570 nm is set to Ch1 and that emitting a longer light wavelength is set to Ch2 automatically.
Page 126 APPLIED OPERATIONS /Image Acquisition From the page tabs on the bottom right of the [Acquire] panel, select the [Settings] sub-panel. <Scope Control> button Displays useful information for the system setup. Fig. 2-4 [Settings] Sub-panel Select the <Scope Control> button. The window as shown below will appear. Fig.
Page 127: Configuring The Scan Unit
APPLIED OPERATIONS /Image Acquisition Select the <BI> button in the [Observation] group box, and select the microscopy from the option buttons below it. The system setting points to be changed for <BI> button microscope observation will blink in red. Change the system configuration (setup of light path selector, etc.) by following the guidance given by the red blinking light.
Page 128: Setting The Observation Condition
APPLIED OPERATIONS /Image Acquisition 2-2-1-3 Setting the Observation Condition 1 Setting the Objective Ma gnification From the drop-down list on the center of the [Acquire] panel, select the objective being used with the microscope. NOTE If the magnification of the objective in use and that set here do not match, the measurement results will be inaccurate.
Page 129 APPLIED OPERATIONS /Image Acquisition To display the information on all channels simultaneously, right-click the boundary between channel display boxes. Click the boundary again to return to the original display. 4 Setting the Highest Scan Speed Set the scan speed to the fastest speed by using the scale in the [Scan Speed] group box in the [Acquire] panel [Scan Speed] group box Set the scan speed by clicking a point on...
Page 130 APPLIED OPERATIONS /Image Acquisition 5 Setting the XY Observat ion Mode In the [Mode] group box, select the [Surface] option button. In the [Acquire] panel, select the XY observation mode option button. 6 Repeated Scanning Ope ration Select the <XY Repeat> button. The acquired image will be displayed in the [Live] panel.
Page 131 APPLIED OPERATIONS /Image Acquisition 7 Setting the Multiple Sec tions to be Observed While acquiring image, move the Z stage to select the multiple sections to be observed. From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Z Stage] sub-panel.
Page 132 APPLIED OPERATIONS /Image Acquisition 8 Setting the Area to be O bserved When the observation targets are concentrated in a narrow area or when it is necessary to observe the detail of a specific area, the image of a limited area can be acquired selectively.
Page 133 APPLIED OPERATIONS /Image Acquisition 9 Adjusting the image brig htness Clicking this button allows fine adjustment of the value. Clicking this field allows the value to be changed on a large scale. The ND value which are usually used are displayed in green. Clicking the <+>...
Page 134 APPLIED OPERATIONS /Image Acquisition NOTE When the light incident to photomultiplier tube is too bright or PMT voltage is set to a high voltage, PMT Over warning may be displayed as shown below. This warning is displayed to protect photomultiplier tube when the light incident to it exceeds a certain level.
Page 135 APPLIED OPERATIONS /Image Acquisition 10 Setting a Lower Scan Sp eed 1. The scan speed can be decreased using the scale in the [Scan Speed] group box on the center of the [Acquire] panel. In general, setting a lower scan speed allows the acquired image quality to be improved.
Page 136: Acquiring Image
APPLIED OPERATIONS /Image Acquisition 11 Stopping Repeated Scan ning After the brightness and gain have been adjusted, select the <STOP SCAN> button in the [Acquire] panel to stop scanning temporarily. 2-2-1-4 Acquiring Image Select the <Once> button. The acquired image will be displayed in the [Live] panel. <Once>...
Page 137 APPLIED OPERATIONS /Image Acquisition 1 Acquiring Image in Accu mulation Mode (Frame mode) From the page tabs on the bottom right of the [Acquire] panel, select the [Settings] sub-panel. [Filter Mode] group box Select the accumulation mode. accumulation modes, [Kalman] [Accumulate To Peak] are available.
Page 138 APPLIED OPERATIONS /Image Acquisition When [Kalman] is selected, enter the accumulation count in the text box. And select the [Frame Kalman] option button displayed. Enter the accumulation count. [Frame Kalman] option button The accumulation count can be set up a maximum of 63 times. When 0 is set as the number of times of accumulation, an ordinary image acquisition is to be performed.
Page 139 APPLIED OPERATIONS /Image Acquisition 2 Acquiring Image in Accu mulation Mode (Line mode) NOTE Image acquisition in the line mode can be performed when you use the FV300 system with AOTF (FV5-COMBA). From the page tabs on the bottom right of the [Acquire] panel, select the [Settings] sub-panel.
Page 140: Saving The Acquired Image In File
APPLIED OPERATIONS /Image Acquisition The accumulation count can be set up a maximum of 63 times. When 0 is set as the number of times of accumulation, an ordinary image acquisition is to be performed. Click the <Once> button in the [Acquire] panel. The acquired image will be displayed in the [Live] panel.
Page 141: Image Acquisition In Other Observation Modes
APPLIED OPERATIONS /Image Acquisition 2-2-2 Image Acquisition in O ther Observation Modes 2-2-2-1 XZ Observation Mode The description in this section will be focused on the image acquisition operations in the XZ observation mode that are not used in the XY observation modes (which are the operations enclosed in in the chart on the next page).
Page 142 APPLIED OPERATIONS /Image Acquisition Set the dyeing method. Set the range of the multiple sections (Section 2-2-1-1 in [OPERATION]) to be observed (the Z-direction scanning range). (Section 2-2-2-1-1 in [OPERATION]) Configure the microscope and scan unit. (Sections 2-2-1-1 & 2-2-1-2 in [OPERATION]) Set a lower scan speed.
Page 143 APPLIED OPERATIONS /Image Acquisition 1 Setting the Z-direction s canning range While acquiring image, move the Z stage according to the range of the multiple sections to be observed (i.e. the Z-direction scanning range). From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Z Stage] sub-panel.
Page 144 APPLIED OPERATIONS /Image Acquisition Do not turn the fine focus adjustment knob while the [Locked] check box is checked, for this may damage the Z motor. 2. While observing the image in the [Live] panel, locate the upper edge of the range to be observed by moving down the stage using the <Z stage coarse adjustment>...
Page 145 APPLIED OPERATIONS /Image Acquisition 2 Setting the observation mode From the page tabs on the bottom right of the [Acquire] panel, select the [Settings] sub-panel. Click the [Depth] option button in the [Mode] group box. In the [Acquire] panel, select the XZ observation mode option button. 3 Setting the observation line A line is displayed on the image in the [Live] panel.
Page 146 APPLIED OPERATIONS /Image Acquisition 5 Acquiring image Click the <XZ> button in the [Acquire] panel. The acquired image will be displayed in the [Live] panel. OPERATION INSTRUCTIONS Page IV . 2 - 3 2...
Page 147: Xt Observation Mode
APPLIED OPERATIONS /Image Acquisition 2-2-2-2 XT Observation Mode The description in this section will be focused on the image acquisition operations in the XT observation mode that are not used in the XY observation modes (which are the operations enclosed in in the chart on the next page).
Page 148 APPLIED OPERATIONS /Image Acquisition 1 Setting the observation mode Click the [Line] option button in the [Mode] group box. In the [Acquire] panel, select the XT observation mode option button. 2 Setting the observation line A line is displayed on the image in the [Live] panel. Place the mouse pointer arrow on the line and drag it to the position you want to observe.
Page 149: Xzt Observation Mode
APPLIED OPERATIONS /Image Acquisition 2-2-2-3 XZT Observation Mode The description in this section will be focused on the image acquisition operations in the XZT observation mode that are not used in the XY observation modes (which are the operations enclosed in in the chart on the next page).
Page 150 APPLIED OPERATIONS /Image Acquisition 1 Setting the Z-direction s canning range While acquiring image, move the Z stage according to the range of the multiple sections to be observed (Z-direction scanning range). From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Z Stage] sub-panel.
Page 151 APPLIED OPERATIONS /Image Acquisition While observing the image in the [Live] panel, locate the upper edge of the range to be observed by moving down the stage using the <Z stage coarse adjustment> and <Z stage fine adjustment> buttons in the [Z Stage] sub-panel. When using the FLUOVIEW system with an inverted microscope, locate the bottom edge of the range to be observed by moving down the revolving nosepiece using the <Z stage coarse adjustment>...
Page 152 APPLIED OPERATIONS /Image Acquisition 4 Setting the numbers of s teps and acquired image slices From the page tabs on the bottom right of the [Acquire] panel, select the [Z Stage] sub-panel. A number of steps is displayed in the [Step Size] text box. This number can be changed using the <...
Page 153 APPLIED OPERATIONS /Image Acquisition 5 Setting the interval time From the page tabs on the bottom right of the [Acquire] panel, select the [Time Series] sub-panel. Set the interval time using the < > or < > button in the [Interval] text box. [Interval] text box Set the interval time using the <...
Page 154 APPLIED OPERATIONS /Image Acquisition 7 Acquiring image Click the <XZT> button in the [Acquire] panel. The acquired image will be displayed in the [Live] panel. While acquiring an image in the XZT observation mode, clicking the <STOP SCAN> button changes the buttons at the upper part of the [Acquire] panel as shown below. The <Resume>...
Page 155: Xyz Observation Mode
APPLIED OPERATIONS /Image Acquisition 2-2-2-4 XYZ Observation Mode The description in this section will be focused on the image acquisition operations in the XYZ observation mode that are not used in the XY observation modes (which are the operations enclosed in in the chart on the next page).
Page 156 APPLIED OPERATIONS /Image Acquisition 1 Setting the Z-direction s canning range While acquiring image, move the Z stage according to the range of the multiple sections to be observed (Z-direction scanning range). From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Z Stage] sub-panel.
Page 157 APPLIED OPERATIONS /Image Acquisition Do not turn the fine focus adjustment knob while the [Locked] check box is checked, for this may damage the Z motor. While observing the image in the [Live] panel, locate the upper edge of the range to be observed by moving down the stage using the <Z stage coarse adjustment>...
Page 158 APPLIED OPERATIONS /Image Acquisition 2 Setting the numbers of s teps and acquired image slices Set the number of steps using the < > or < > button in the [Step Size] text box. The number of steps shown in the [Step Size] text box has been calculated [Step Size] text box by the system so that the depth scale of the acquired image is identical to the horizontal scale.
Page 159 APPLIED OPERATIONS /Image Acquisition 5 Appending image XZT image can be added after the image acquisition. Immediately after acquisition of an image in the XYZ observation mode, the buttons at the upper part of the [Acquire] panel changes as shown below. <Series Done>...
Page 160: Xyt Observation Mode
APPLIED OPERATIONS /Image Acquisition 2-2-2-5 XYT Observation Mode The description in this section will be focused on the image acquisition operations in the XYT observation mode that are not used in the XY observation modes (which are the operations enclosed in in the chart on the next page).
Page 161 APPLIED OPERATIONS /Image Acquisition 1 Setting the observation mode Click the [Surface] option button in the [Mode] group box in the [Acquire] panel. In the [Acquire] panel, select the XYT observation mode option button. 2 Setting the interval time From the page tabs on the bottom right of the [Acquire] panel, select the [Time Series] sub-panel.
Page 162 APPLIED OPERATIONS /Image Acquisition 3 Setting the number of sc ans Set the number of scans using the < > or < > button in the [N] text box in the [Time Series] sub-panel. 4 Acquiring image Click the <XYT> button in the [Acquire] panel. The acquired image will be displayed in the [Live] panel.
Page 163: Xyzt Observation Mode
APPLIED OPERATIONS /Image Acquisition 2-2-2-6 XYZT Observation Mode The description in this section will be focused on the image acquisition operations in the XYZT observation mode that are not used in the XY observation modes (which are the operations enclosed in in the chart on the next page).
Page 164 APPLIED OPERATIONS /Image Acquisition Set the dyeing method. (Section 2-2-1-1 in [OPERATION]) Set the range of the multiple Configure the microscope and scan unit. sections to be observed (the Z- (Sections 2-2-1-1 & 2-2-1-2 in [OPERATION]) direction scanning range). (Section 2-2-2-6-1 in [OPERATION]) Set the objective magnification.
Page 165 APPLIED OPERATIONS /Image Acquisition 1 Setting the Z-direction s canning range While acquiring image, move the Z stage according to the range of the multiple sections to be observed (Z-direction scanning range). From the panel page tabs shown on the bottom right of the [Acquire] panel, select the [Z Stage] sub-panel.
Page 166 APPLIED OPERATIONS /Image Acquisition Do not turn the fine focus adjustment knob while the [Locked] check box is checked, for this may damage the Z motor. While observing the image in the [Live] panel, locate the upper edge of the range to be observed by moving down the stage using the <Z stage coarse adjustment>...
Page 167 APPLIED OPERATIONS /Image Acquisition 2 Setting the numbers of s teps and acquired image slices Set the number of steps using the < > or < > button in the [Step Size] text box. The number of steps shown in the [Step Size] text box has been calculated [Step Size] text box by the system so that the depth scale of the acquired image is identical to the horizontal scale.
Page 168 APPLIED OPERATIONS /Image Acquisition 4 Setting the interval time From the page tabs on the bottom right of the [Acquire] panel, select the [Time Series] sub-panel. A panel as shown below will be displayed. [Interval] text box Set the interval time using the < >...
Page 169 APPLIED OPERATIONS /Image Acquisition 6 Acquiring image Click the <XYZT> button in the [Acquire] panel. The acquired image will be displayed in the [Live] panel. When the built-in power supply for transmitted light illumination is used for a long period in the XYZT mode, the metallic parts may be expanded by heat, causing the focusing to be deviated.
Page 170: Differences In Image Acquisition Method Between Fluorescent And Transmitted Images
APPLIED OPERATIONS /Image Acquisition 2-2-3 Differences in Image A cquisition Method Between Fluorescent and Transmitted Images 2-2-3-1 Monochrome Image The wavelength obtained by monochrome dyeing can be acquired and observed as an image of a channel (either Ch1 or Ch2). The subsequent description deals with the differences in operation between the monochrome dyeing and dual-fluorochrome dyeing.
Page 171 APPLIED OPERATIONS /Image Acquisition Set the dyeing method From the page tabs on the bottom right of the [Acquire] panel, select the [Dyes] sub-panel. Place the pointer on the icon displayed [Assign dyes manually] check box [Selected Dyes], and the Checking this enables the manual dyeing method is shown in setting.
Page 172 APPLIED OPERATIONS /Image Acquisition One Point! The [Assign dyes manually] check box can also be used to set the dyeing method to the desired channel. Check the [Assign dyes manually] check box in the [Dyes] sub-panel. Select the dyeing method in the [Available Dyes] list box and drag it directly to the field of the [Ch] check box.
Page 173 APPLIED OPERATIONS /Image Acquisition Make the channel ready for image acquisition. In the [Acquire] panel, make sure that the check box in the [Ch1] or [Ch2] group box that shows the dyeing method is check-marked to indicate that the corresponding channel is ready for image acquisition.
Page 174: Dual-Fluorochrome Image
APPLIED OPERATIONS /Image Acquisition 2-2-3-2 Dual-Fluorochrome Ima ge The wavelength obtained by dual-fluorochrome dyeing can be acquired and observed as images of the channels (Ch1 or Ch2). The subsequent description deals with the differences in operation between the monochrome dyeing and dual-fluorochrome dyeing. When the specimen dyed by dual-fluorochrome dyeing is observed, the image of wavelengths below 570 nm is acquired in channel 1 (Ch1) and the image of wavelengths above 570 nm is acquired in channel 2 (Ch2).
Page 175 APPLIED OPERATIONS /Image Acquisition When dyeing methods are selected from the [Available Dyes] list box and the <Apply> button is clicked to apply them in the image acquisition channel, the dyeing methods emitting a shorter light wavelength than 570 nm are set to Ch1 and those emitting a longer light wavelength are set to Ch2 automatically.
Page 176 APPLIED OPERATIONS /Image Acquisition Engage the filter by setting the [BARRIER FILTERS] lever of the scan unit. (Setting the lever to “IN” engages the filter.) Also see section 2-2-1-2, “Configuring the Scan Unit” and follow the instructions given by the [Microscope Configuration] window. Make the two channels ready for image acquisition.
Page 177: Transmitted Image
APPLIED OPERATIONS /Image Acquisition 2-2-3-3 Transmitted Image Images obtained by transmitted light observation can also be acquired or observed simultaneously with images obtained by fluorescence observation. Set the [DETECTION MODE] lever of the scan unit as shown below. FL -: Fluorescence observation with monochrome or dual-fluorochrome dyeing (Ch1), and transmitted light observation (Ch3).
Page 178 APPLIED OPERATIONS /Image Acquisition When dyeing methods are selected from the [Available Dyes] list box and the <Apply> button is clicked to apply them in the image acquisition channel, the dyeing methods emitting a shorter light wavelength than 570 nm are set to Ch1 and those emitting a longer light wavelength are set to Ch2 automatically.
Page 179 APPLIED OPERATIONS /Image Acquisition Engage the filter by setting the [BARRIER FILTERS] lever of the scan unit. (Setting the lever to “IN” engages the filter.) Also see section 2-2-1-2, “Configuring the Scan Unit” and follow the instructions given by the [Microscope Configuration] window. Turn off the transmitted light bulb of the microscope.
Page 180 APPLIED OPERATIONS /Image Acquisition Make channel ready for transmitted image acquisition. In the [Acquire] panel, make sure that the [Transmitted] check box in the [Ch3] group box is check-marked to indicate that Ch3 is ready for image acquisition. Ch3 check box When observing a fluorescence image simultaneously, set the required channel ready for acquisition of fluorescence image.
Page 181: Image Acquisition By Rotating It (Rotation Scan)
APPLIED OPERATIONS /Image Acquisition 2-2-4 Image Acquisition by Rotating It (Rotation Scan) Rotation scan enables image acquisition with tilting the field of viewed. NOTE Rotation Scan can not be used with Free Line Scan. This function is available when the FV5-IO3 is in use. NOTE NOTE If you do not use the FV5-IO3, the [Pan] group box does not display the...
Page 182: Image Acquisition Of Only The Rectangular Position (Clip Scan)
APPLIED OPERATIONS /Image Acquisition 2-2-5 Image Acquisition of O nly the Rectangular Position (Clip Scan) The Clip Scan mode limits the image acquisition range to the range to be observed and acquires the image of only that range. Applying this mode in an image acquisition mode involving large amount of data, such as the XYZ, XYT and XYZT observation modes, makes it possible to acquire data of a long period of time in a small file size.
Page 183 APPLIED OPERATIONS /Image Acquisition Change the frame size. To change the frame size, click a point inside the frame with the mouse pointer. When square handles are displayed on the frame edges, place the mouse pointer on one of them and drag it. Handle Change the inclination angle of the line.
Page 184 APPLIED OPERATIONS /Image Acquisition It is not permitted to change the scanning range after having acquired NOTE images. If you want to change the scanning range again, select <Normal> from the list displayed below the [Surface XY-Norm] option button in the [Mode] group box to set the scanning ragne to the original setting, acquire an image in the XY observation mode, then select <Clip>...
Page 185 APPLIED OPERATIONS /Image Acquisition Acquire an image in the XY observation mode. For the operation procedure, see section 2-2-1, “Image Acquisition with XY observation”. Select <ZoomIn> under the [Surface XY] option button in the [Mode] group box in the [Acquire] panel. A frame indicating the control range appears in the [Live] panel.
Page 186 APPLIED OPERATIONS /Image Acquisition Change the inclination angle of the frame. Click the mouse right button inside the frame and, when a pop-up menu appears, select [Rotate] from it. The inclination angle of the frame can be varied according to the mouse pointer movement.
Page 187: High-Speed Image Acquisition
APPLIED OPERATIONS /Image Acquisition 2-2-7 High-Speed Image Ac quisition An image can be acquired in 0.25 second. This image acquisition mode is valid under the following condition: Max. 2 channels Image size: 512 x 512 pixels Zoom ratio of 2X or more, with setting in 2X steps Check one or both of the [Ch] check boxes in the [Ch] group box in the [Acquire] panel.
Page 188: Image Acquisition To Prevent Crosstalk Between Fluorescence
APPLIED OPERATIONS /Image Acquisition 2-2-8 Image acquisition to p revent crosstalk between fluorescence (Sequential Scan) An image which is suppressed crosstalk can be acquired sequentially by combination of excitation laser and those image acquisition channel. With this image capturing method, the image of a multiple-dyed specimen can be obtained by sequentially acquiring image slice of each type of fluorescence.
Page 189 APPLIED OPERATIONS /Image Acquisition [Group] (which means each laser) and the < > and < > buttons appear on the lower part of each [CH] group box. Click to increase the acquired group Shows the group to Click to decrease be acquired.
Page 190 APPLIED OPERATIONS /Image Acquisition <Off> button Select the laser type. Fig. 2-23 [Lasers] Sub-panel Among the laser <Off> buttons on the right of the [Laser Intensity] scales in the [Lasers] group box, click the <Off> button of the laser you want to use in transmitted observation.
Page 191: Virtual Channel Function
APPLIED OPERATIONS /Image Acquisition 2-2-8-1 Virtual Channel Functio n The virtual channels refer to the simulated detection channels produced when a single detector switched in sequential scan. The virtual channels make it possible to perform sequential scan by switching the lasers and filters according to the preset condition by increasing the number of detection channels.
Page 192 APPLIED OPERATIONS /Image Acquisition In the [Mode] group box in the [Acquire] panel, click the [Surface XY-Norm] option button, select <Sequen> from the list below it and select the scan mode to be used from the displayed icons. Do not select <Line Aq> in the [Mode] group box in the [Acquire] panel NOTE because the virtual channels are not compatible with Line Sequential Scan.
Page 193 APPLIED OPERATIONS /Image Acquisition A virtual channel will be added in the [Ch] group box. (Example: When [Using PMT 1] and [Using PMT 2] are selected from the pop-up menus.) Virtual channel NOTE It is not permitted to set the physically identical channels (an ordinary channel and virtual channel) in the same group.
Page 194 APPLIED OPERATIONS /Image Acquisition Set the laser and filter types condition. Select <SU Control> button of the [Settings] sub-panel in the [Acquire] panel to display the [Optical System Configuration] window. In the [Optical System Configuration] window, set the dyeing method of virtual channel, then set the displayed optimum filter, etc.
Page 195 APPLIED OPERATIONS /Image Acquisition NOTE Filters cannot be set if you are using the FV300. Please switch manually. NOTE Setting will disappear if <Normal> is selected at [Mode] group box of [Acquire] panel after setting virtual channel. It is useful that the setting is saved with use of <Save>...
Page 196: Image Acquisition Of A Line At Desired Angle
APPLIED OPERATIONS /Image Acquisition Click the <STOP SCAN> button to stop repeated scanning. Repeat steps 7 and 10 for each of the groups to be configured. Click the <Seq. Once> button to start sequential scan. <Seq Once> button When a group switches, the following dialog box is displayed. Set filter types etc. in the scanning unit and selects <O.K.>...
Page 197 APPLIED OPERATIONS /Image Acquisition Acquire an image in the XY observation mode. For the operation procedure, see section 2-2-1, “Image Acquisition with XY observation”. Select [Line XT] option button or <Slant> under the [Depth XZ] option button in the [Mode] group box in the [Acquire] panel. A line indicating the control range appears in the [Live] panel.
Page 198 APPLIED OPERATIONS /Image Acquisition The inclination angle of the line can be varied according to the mouse pointer movement. Place the mouse pointer on an inclined position and click the mouse left button to fix the line inclination angle. Set the interval time or the number of scans. The operation procedure is same as that in the XY observation mode.
Page 199: Display The Change Of Image Intensity (Point Scan)
APPLIED OPERATIONS /Image Acquisition 2-2-10 Display the change of image intensity (Point Scan) The change of fluorescence intensity according to time-lapse can be displayed graphically by irradiating the laser to certain point on the image. If you have FLUOVIEW TIME COURSE software, image acquisition can be started with buttons, keys or input of external trigger signals.
Page 200 APPLIED OPERATIONS /Image Acquisition The graph window showing the cross cursor for scanning and the intensity values appears in the [Live] panel. Cross cursor Graph window Move the cross cursor to the area you want to observe. To move the cross cursor, place the mouse pointer on it and drag the mouse.
Page 201 APPLIED OPERATIONS /Image Acquisition The speed recommended for point scan are as follows. 2 s(The scroll bar indicates “Fast” position. It is useful for the specimen changes rapidly.), 10 s, 100 s(The scroll bar indicates “Slow” position. It is useful for the specimen changes slowly.) Set the time for measurement.
Page 202 APPLIED OPERATIONS /Image Acquisition The image shown in the [Live] panel after image acquisition is the image that acquired at the coordinates specified in the [Live] panel and arranged in the X direction, from the top left to the bottom right. The image shown after image acquisition is the same a width as that in the X direction specified in the [Live] panel.
Page 203: Image Acquisition On Desired Line (Xz, Xt Or Xzt Observation)
APPLIED OPERATIONS /Image Acquisition When click the mouse on the X or Y Axis on the graph window, the [Editing] dialog box appears and the graph parameters or the graph display method can be modified. After image acquisition, select <Normal> under the [Surface XY] option button in the [Mode] group box in the [Acquire] panel to return from the image acquisition mode.
Page 204 APPLIED OPERATIONS /Image Acquisition Change the curved line shape. Place the mouse pointer within the box and right click to display the pop-up menu. Then select <Edit>. The cross shaped handles appear around the curve. Place the mouse pointer on either handle and drag it to change the line shape.
Page 205 APPLIED OPERATIONS /Image Acquisition Set the interval time or the number of scans. The operating procedure is same as that in the XY observation mode. See section 2-2-2, “XT Observation Mode” for details. Up to 8000 times, images can be acquired. Set the number of scans in the [N] text box in the [Time Series] sub-panel.
Page 206: Image Acquisition In The Laser Excitation Mode
APPLIED OPERATIONS /Image Acquisition 2-2-12 Image Acquisition in t he Laser Excitation Mode When you use the FV system with AOTF (FV5-COMBA), the function described in this section is available. In the FV system with AOTF, it is possible to cut excitation of laser except the region where scanning is performed.
Page 207: Making Rex Mask File
APPLIED OPERATIONS /Image Acquisition In the REX and Bleach mode, image acquisition in the Focus mode, NOTE fast scan, and image acquisition in the Line mode (Normal, Slant, and Free) can not be performed. NOTE The specimen in the images carried in this section is not for FRAP, but for the user’s manual.
Page 208 APPLIED OPERATIONS /Image Acquisition In the list of buttons displayed as shown below, select the <Rectangular>, <Circle>, <Polyregion>, or <Free region> button. And drag on the image to specify the region of laser excitation. <Rectangular> button <Circle> button <Polyregion> button <Free Region>...
Page 209 APPLIED OPERATIONS /Image Acquisition From the page tabs on the bottom right of the [Acquire] panel, select the [Lasers] sub-panel. <Make REX mask> button Makes the REX mask file of the specified region. Fig. 2-24 [Lasers] sub-panel When the <Make REX mask> button is displayed in gray, click and select the specified region on the [Live] panel.
Page 210 APPLIED OPERATIONS /Image Acquisition Select the <Make REX mask> button. <Make REX mask> button The [REX Live] panel (the REX mask file) is made in the [Display] panel. Make the REX mask file which masks the region outside of the specified region.
Page 211 APPLIED OPERATIONS /Image Acquisition In order to adjust the laser intensity for every region, right-click the mouse on the white region on the [Display] panel where the REX mask file is displayed. The pop- up menu as shown below appears. Select [Set intensity] in the menu.
Page 212: Example Of Frap Experiment
APPLIED OPERATIONS /Image Acquisition 2-2-12-2 Example of FRAP exper iment The example of procedure of FRAP experiment using AOTF is described in this section. In order to perform photobleaching to a specimen, the value of laser intensity is raised to 100% to acquire an image since strong laser intensity is temporarily required.
Page 213 APPLIED OPERATIONS /Image Acquisition Select the <Once> button in the [Acquire] panel and acquire an image. If necessary, set the range for image acquisition with clip scan, for example, after acquiring an image. 3 Making REX Mask File Refer to section 2-2-12-1, “Making REX Mask File” for the procedure to make the REX mask file.
Page 214 APPLIED OPERATIONS /Image Acquisition 4 Selecting REX Mask File and Setting the Laser ON/OFF Set the REX mask file in the REX mode and Bleach mode, and the laser ON/OFF respectively. Select the [REX mode] option button in the [AOTF] group box in the [Lasers] sub- panel.
Page 215 APPLIED OPERATIONS /Image Acquisition Select the [Bleach mode] option button in the [AOTF] group box of the [Laser] sub- panel. A frame to specify the REX mask file appears on the right side of the [Bleach mode] option button. Right-click the mouse inside the frame to display the pop-up menu as shown below.
Page 216 APPLIED OPERATIONS /Image Acquisition 5 Setting the XYT Observa tion mode Confirm that [XY-Norm] is displayed in the [Mode] group box in the [Acquire] panel. Select the [XYT] option button in the [Acquire] panel. OPERATION INSTRUCTIONS Page IV . 2 - 1 0 2...
Page 217 APPLIED OPERATIONS /Image Acquisition From the page tabs on the bottom right of the [Acquire] panel, select the [Time Series] sub-panel. The panel as shown below appears. [Interval] text box Set the interval with the < > and< > buttons. Key entry is also acceptable. [N] text box Set the number of scans with the <...
Page 218 APPLIED OPERATIONS /Image Acquisition 7 Acquiring Data The experiment data before and after photobleaching of certain region can be obtained after performing an ordinary image acquisition in the first scan, an image acquisition in the Bleach mode in the second scan , and an ordinary image acquisition in the third scan.
Page 219 APPLIED OPERATIONS /Image Acquisition Start the second scan in 10 seconds after the first scan is performed. The laser of strong intensity is irradiated specified region. Select the [Disabled] or [REX mode] option button in the [AOTF] group box in the [Lasers] sub-panel.
Page 220 APPLIED OPERATIONS /Image Acquisition Start the third scan in 10 seconds after the second scan is performed. The image of the specimen is fading only where the laser is irradiated. (The [Disabled] option button is selected.) Start the forth and subsequent scan in 10 seconds after the third scan is performed.
Page 221 APPLIED OPERATIONS /Image Acquisition 2-2-13 Notes for image acqui sition 2-2-13-1 Memory of setting inform ation for scanning region Frame with handle described in 2-2-5 Clip Scan, Frame with hand described in 2-2-6 Zoom-in Scan, Straight line with handle described in 2-2-9, Cross line cursor described in 2-2-10 Point Scan are all called Scanning Region Setting - ROI (Region Of Interest).
Page 222 APPLIED OPERATIONS /Image Acquisition Kind of I/O Observ Acquiring mode board ation I/O2 I/O3 Normal scan Clip scan Zoom-in scan Fast normal scan Fast clip scan Sequential normal scan Sequential clip scan Sequential zoom-in scan Line sequential normal scan Line sequential clip scan Line sequential zoom-in scan Normal scan Clip scan...
Page 223 APPLIED OPERATIONS /Image Acquisition Kind of I/O board Mode Sub Mode I/O2 I/O3 XYZT Normal scan Clip scan Zoom-in scan Sequential normal scan Sequential clip scan Sequential zoom-in scan Line sequential normal scan Line sequential clip scan Line sequential zoom-in scan Normal scan Clip scan Zoom-in scan...
Page 224 APPLIED OPERATIONS /Image Acquisition It is possible to identify which I/O board is used with the following method. In case of I/O2, the arrow of [Pan][Zoom] group box of [Acquire] panel is grayed out and cannot be selected when it boots up as shown below. It is grayed out.
Page 225: Saving, Opening And Shredding Images
APPLIED OPERATIONS /Saving, Opening and Shredding Images 2-3 Saving, Opening and Shredding Images Use the [File I/O] panel to save, open or shred an image. Display the [File I/O] panel at the front. [Open] [Display] panel To open a file, select the desired file in the [Files] list box and drag the file here.
Page 226 APPLIED OPERATIONS/ Saving, Opening and Shredding Images <Image Icons> Images are represented by icons which can also Image Icon Significance identify the observation modes used when acquiring XZ observation them. XZ observation, 2-channel mode The icon of the selected image (image in the [Display] panel) is displayed in the frame at the top Xt observation of the function panel such as the [File I/O] panel.
Page 227 APPLIED OPERATIONS/ Saving, Opening and Shredding Images <[Files] List Box> The information displayed in the list box can be changed. <Details> button To display all information, click the <Details> button above the [Files] list box to broaden it. <List> button To return to the display of icons and file names only, click the <List>...
Page 228: Saving Images
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-1 Saving Images Either a series of images or single image being displayed can be saved. 2-3-1-1 Saving Images As a Series The displayed images can be saved in a disk as a series image file. Display the [Display] panel showing the image to be saved at the front.
Page 229 APPLIED OPERATIONS/ Saving, Opening and Shredding Images When it is required to change the save destination drive or directory, use the [Save in:] drop-down list. When it is required to change the saved file type, use the [Save as Type:] drop- down list.
Page 230: Saving A Display
APPLIED OPERATIONS/ Saving, Opening and Shredding Images One Point! Live image file is stored to the assigned folder with NOTE [Seq. Number] function is enabled when [Save Experiment As] dialog box appears for not stored Live image is existing. Seq. Numbering function is disabled once after the [Save Experiment As] setting is changed.
Page 231 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Fig. 2-29 [Save Experiment As] Dialog Box OPERATION INSTRUCTIONS Page IV . 2 - 1 1 7...
Page 232 APPLIED OPERATIONS/ Saving, Opening and Shredding Images When it is required to change the save destination drive or directory, use the [Save in:] drop-down list. When it is required to change the saved file type, use the [Save as Type:] drop- down list.
Page 233: Saving Specified Area Of Image
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-1-3 Saving Specified Area of Image Only the specified part of an image can be saved as an image in a disk. Display the [Display] panel of the image containing the part to be saved in the front. When the image was acquired through multiple channels, you can select whether the image slices for multiple channels are saved together or the image slice of only 1 channel is saved.
Page 234 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Specify the area to be saved in the image in the [Display] panel. The area is displayed on the image with the handles on its frame. The area becomes the save target while these handles are displayed.
Page 235 APPLIED OPERATIONS/ Saving, Opening and Shredding Images 12. A dialog box as shown below appears. Size of the specified area Fig. 2-31 [Saving a subset of Experiment’s Data]Dialog Box The [Saving a subset of Experiment’s Data] dialog box does not appear if no area is specified in the image on the [Display] panel.
Page 236: Saving Animation Images
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-1-4 Saving Animation images Transform a created animation image into the AVI file format and save it in the file type which can display the animation image without this software. Create an animation image. Refer to “2-9-2 Animation”...
Page 237 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Click the <Animation> button in the [Save] group box. The [Save Experiment As] <Save Animation> button dialog box appears as shown below. Fig. 2-32 [Save Experiment As] Dialog Box When it is required to change the save destination drive or directory, use the [Save in:] drop-down list.
Page 238: File Types Available For Save
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-1-5 File Types Available for Save The file type used for saving image in a file can be selected by the user. See section 2- 3-1-1, “Saving Images As a Series” for the operation method. Click the <Experiment>...
Page 239 APPLIED OPERATIONS/ Saving, Opening and Shredding Images NOTE If you want to save the image with the comment drawn on it as an image, select the file type according to the usage of the image as described below. Fluoview Multi Tiff:Select when the image will be analyzed or processed after save.
Page 246: Opening Previously Saved Images
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-2 Opening Previously Saved Images Image files saved in the disk can be opened as follows. If the image file name that you want to open is not displayed in the [Files] list box, change the drive and/or directory to those containing the desired file using the [Drive] drop-down list and/or [Directory] list box.
Page 247: Shredding Images
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-3 Shredding Images Shredding an image refers to removing it from the objects of processing including display. Shredding does not actually deletes the image saved in the disk. Click the <Experiment List> button in the toolbar at the bottom of the [File I/O] panel.
Page 248 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Multiple Images displayed can be shredded simultaneously. Display the [Experiments in Memory] dialog box while showing multiple images in the [Display] panel. Pressing down the Shift key, select multiple image files to be shredded.
Page 249: Saving Comment Together With Image
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-4 Saving Comment Together with Image Click the <Experiment List> button in the toolbar at the bottom of the [File I/O] panel. The [Experiments in Memory] dialog box appears as shown below. <Experiment List> button Fig.
Page 250 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Enter comment from the keyboard. NOTE During modification of previously entered comment, if it is required to restore the originally entered comment, click the <Restore Comments> button. However, once the <Save Comments To Image> button is pressed, the original comment cannot be restored.
Page 251: Checking The Image Information/Acquisition Parameters
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-5 Checking the Image Information/Acquisition Parameters Click the <Experiment List> button in the toolbar at the bottom of the [File I/O] panel. The [Experiments in Memory] dialog box appears as shown below. <Experiment List> button Fig.
Page 252 APPLIED OPERATIONS/ Saving, Opening and Shredding Images The [X Size], [Y Size] and [Z Size] text boxes can also be used to change the lengths per image pixel or the number of steps in the Z-direction. These values are used in the scale display and many other analysis operations. When opening and analyzing an image file creased with another application on FLUOVIEW, enter the lengths per image pixel and the number of steps in the Z-direction if these values are known, then click the...
Page 253 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Display the [Image Fields] panel at the front. The panel as shown below appears, where the information on the selected image is shown. Range of intensity values mapped for each channel. LUT status. Acquisition parameters.
Page 254 APPLIED OPERATIONS/ Saving, Opening and Shredding Images One Point! The [Image Comments] dialog box can also be displayed by a mouse operation. Display the image to be saved at the front of the [Display] panel, and right- click a point in the image. A pop-up menu as shown below is displayed.
Page 255: Saving The Image Information/Observation Condition
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-6 Saving the Image Information/Observation Condition The image information and observation condition can be saved as ASCII text file in a disk. Display the [Display] panel of the image to be saved at the front position. Select the <Experiment List>...
Page 256 APPLIED OPERATIONS/ Saving, Opening and Shredding Images In the list in the [Experiments in Memory] dialog box, select the file name including the image whose image information and observation condition are to be saved. And click the <Comments> button. The [Image Comments] dialog box as shown below appears.
Page 257 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Select the <Save> button. The [Save As ASCII Text] dialog box as shown below appears. Fig. 2-42 [Save As ASCII Text] dialog box In order to change the save destination drive or directory, use the [Save in: ] drop- down list.
Page 258: Saving/Reading The Region File
APPLIED OPERATIONS/ Saving, Opening and Shredding Images 2-3-7 Saving/Reading the Region File The information on the shape, position, and color of certain region can be saved in a file and they can also be read. 2-3-7-1 Saving the Region File Display the [Display] panel of the image including the region to be saved at the front position.
Page 259 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Select the image file name including the region to be saved in the [Experiments in Memory] dialog box and click the <Comments> button. The [Image Comments] dialog box as shown below appears. <Annotations/ROIs> button in the [Export to File] group Saves the region into a file.
Page 260: Reading The Region File
APPLIED OPERATIONS/ Saving, Opening and Shredding Images Enter the file name into the [File name] text box and click the <Save> button. When the text file having the same name as the entered name is NOTE already exists, the dialog box appears to ask you to overwrite the existing file or not.
Page 261 APPLIED OPERATIONS/ Saving, Opening and Shredding Images <Annotations/ROIs> button in the [Export to File] group Saves the region into a file. <Annotations/ROIs> button in the [Import from File] group Reads the file saving the region. Fig. 2-47 [Image Comments] dialog box Select the <Annotations/ROIs>...
Page 262 APPLIED OPERATIONS/ Saving, Opening and Shredding Images Select the file to be read and click the <Open> button. When the image size of the file including the region to be read and that of the specified file differ, the [Careful] dialog box as shown below appears. Clicking the <OK>...
Page 263: Protocol Processor
APPLIED OPRETIONS /protocol processor 2-4 Protocol processor The Protocol Processor makes FLUOVIEW image acquisition flexible, especially time series image acquisition. The function enables to create and modify the experiment procedure such as image acquisition condition with interval setting, laser excitation area and its power setting. The protocol can save and modify to be suitable to new experiment.
Page 264: Starting The Protocol Processor
APPLIED OPERATIONS / Protocol processor The protocol can be edited by Protocol Processor software that comes with the FLUOVIEW software. 2-4-1 Starting the Protocol Processor Starts the Protocol Processor by clicking <Protocol Processor> button in [Time Series] sub-panel of [Acquire] panel. <Protocol Processor>...
Page 265: Editing The Protocol
APPLIED OPERATIONS /Protocol processor : <Get FV Status> button: Acquires each setting value of FLUOVIEW software parameters and registers to PAPP window. The track which the parameters to be registered has to be highlighted. : <Set to FV> button: Sets PAPP parameters to FLUOVIEW software parameters..
Page 266: Description Of Setting Items
APPLIED OPERATIONS / Protocol processor 1 Description of Setting Items The [PAPP] window has the setting items as described in the following. <Path> button [Image File Name] text box Specify the folder for <Get Init Values> button Enter the file name in which the image saving image acquired will be saved.
Page 267 APPLIED OPERATIONS /Protocol processor Only available Laser type and PMT channel column is used with [Laser Intensity] and [PMT Voltage]. The values in each cell are applied to the [Acquire] panel at the beginning of the protocol. When “Init” is specified in a cell, the value set in the [Init Val] box is applied. The blank cell applies the value set to the track immediately above.
Page 268 APPLIED OPERATIONS / Protocol processor Normal : Starts image acquisition when protocol comes the Track. Trigger : Starts image acquisition after receiving external trigger input. Wait : Starts image acquisition after wait time describes the track, the unit is second. Condition: See the following table in case that [New mode] or [Series mode] is selected in [Mode].
Page 269 APPLIED OPERATIONS /Protocol processor Pan Status: Enter Pan value, refer to [Pan] [Zoom]group box in [Acquire] panel. Zoom X Pan, Y Pan Angle FLUOVIEW software [Pan][Zoom] Group Box Pan value can not enter the following condition. · Mode is set to any of XY, XYT, XYZ or XYZT. ·...
Page 270 APPLIED OPERATIONS / Protocol processor mode setting. In this case of Normal scan, there might be scanned before the scan mode changes. Scan Status: Change scanning position and its size with the selected scan mode. See 2-4-10 “Restrictions at [PAPP] window input time” for restriction of Scan status input.
Page 271 APPLIED OPERATIONS /Protocol processor mask which is displayed on FLUOVIEW software or to disable the mask by selecting “None”. [REX Mask] appears only when AOTF (FV5-COMBA) is equipped. PMT Voltage: Enter the PMT voltage of each channel. The setting is 1 to 1100, and the unit is V.
Page 272 APPLIED OPERATIONS / Protocol processor One Point! It is possible to designate folder in [Save File Name] column. For example, if [R001 Image001] is entered, folder – [R001] is created under path of [Image File Name:] and image file is saved under the name of [Image001]. Trigger Out: Select the trigger output from “00”, “01”, “10”...
Page 273: Command List
APPLIED OPERATIONS /Protocol processor 2 Command List The list of macro commands is shown below. When “Command” is selected in column [Mode] in the [PAPP] window, enter the macro command to be carried out for the track. The macro command to be entered can be selected from the drop-down list.
Page 274 APPLIED OPERATIONS / Protocol processor case that Invalid is to be set, select OFF. <E.g> In case that channel 1 is to be Valid, enter [EnCh1, ON]. Filter [NORMAL Filter [NORMAL/KALMAN/PEAK]: It defines type of /KALMAN/PEAK] integration. [ ,value] NORMAL: No integration KALMAN: KALMAN filter PEAK: Peak detecting integration value: In case that KALMAN is selected,...
Page 275 APPLIED OPERATIONS /Protocol processor Offset ch value Offset Sets the Offset value of channel. <E.g.> Enter “Offset 2, 5” to set 5 to the offset value of channel 2. PasteROI PasteROI Pastes ROI to the next track. Note: Before the paste, Normal scan is executed one time.
Page 276 APPLIED OPERATIONS / Protocol processor (1024, 1024) (2048, 2048). Stop&Save Stop&Save Saves all the images acquired before the Stop&Save command is executed. Note: The images are saved separately from the images acquired in succeeding tracks. Store Store Opens a new [Display] panel to display the image.
Page 277 APPLIED OPERATIONS /Protocol processor The destination of file save using the Save and Stop&Save commands is the path displayed on the left of the [Image File Name] text box. If it is required to change the file save destination, click the <Path> button on the right of the text box and specify the folder in the displayed dialog box.
Page 278: Protocol Repetition Processing
APPLIED OPERATIONS / Protocol processor 3 Protocol Repetition Processing A protocol can be executed repeatedly for the specified number of times using a variable. Use the For and Next commands to execute repetition. The track enclosed between For and Next can be repeated for the number of times specified using a variable.
Page 279: Input Supporting Function
APPLIED OPERATIONS /Protocol processor One Point! It is possible to use variables in [For] statement in other cell of protocol processor. <E.g> It is possible to change laser intensity value every image acquiring time. When laser intensity of argon laser is defined as variable N, it creates protocols that acquire image with interval of step 10 by N equal to 10 to 100.
Page 280 APPLIED OPERATIONS / Protocol processor All of parameters that can be set for the Track can modify. Parameter that is deferent with the previous Track is colored with Red. Parameters that is modified on [Track] windows is colored with purple. When button is clicked, the setting of item checked at check box of group box name is changed to the value in [Init Val] of [PAPP] window and [init] is...
Page 281: Protocol Procedure Supporting Function
APPLIED OPERATIONS /Protocol processor FLUOVIEW software NOTE setting. [Track] parameter can not modify nor register during the Track is NOTE running. 5 Protocol procedure supporting function [Protocol Creator] window helps to make procedure, especially time-series, easily.. Track No. Drag & Drop Interval time for each track End time of each track Setting value display text box...
Page 282: Saving The Protocol
APPLIED OPERATIONS / Protocol processor [Track] window opens by double-clicking icon on [Track Image] group for detail setting. Details of parameters are displayed in setting value display text box by clicking icon. Interval time of each Track can be entered into the text box. The parameters affect to the [PAPP] window by pressing <Close>...
Page 283 APPLIED OPERATIONS /Protocol processor list. Enter the file name in the [File Name:] text box. Select file type, .pap or .csv, with [Save as type:] dropdown list. Click the <Save> button. NOTE If a CSV file having the same file name as the file name entered above already exists, a dialog box appears to ask if you want to overwrite the existing file.
Page 284: Executing The Protocol
APPLIED OPERATIONS / Protocol processor 2-4-4 Executing the Protocol The created protocol can be executed to acquire time series images. In the [PAPP] window, click the <Start> button. <Start> button The number of rows in the [No.] list in the [PAPP] window becomes 2, and the window display is reduced.
Page 285 APPLIED OPERATIONS /Protocol processor To stop the protocol without executing it till the end, click the <End> button. <Break> button When clicked while the protocol is executed, the <End> button performs the same function as the <Series Done> button in the [Acquire] panel.
Page 286: Loading A Protocol
APPLIED OPERATIONS / Protocol processor 2-4-5 Loading a Protocol The Protocol Processor creates and saves protocol files in the CSV file format. The following procedure loads a CSV file (extension .pap or .csv) for editing the protocol in it. Start the Protocol Processor software and display the [PAPP] window. For details, see section 2-4-1, “Starting the Protocol Processor”.
Page 287: Loading A Protocol Of Previous Format
APPLIED OPERATIONS /Protocol processor 2-4-6 Loading a protocol of previous format FLUOVIEW software Ver4.2 inform the protocol data conversion when the software reads previous version of the data. The rule of the conversion is explained below. Rule A Before Ver4.2, Frame = XYT or XYZT is converted to XYT or XYZT in count of Z slice parameter.
Page 288 APPLIED OPERATIONS / Protocol processor Fig. 2-52 Multi-Point Time Lapse Protocol Rule C For and Command in Series appears with indented + ( ). Combined For – Next structure becomes single layer. Fig. 2-53 Protocol Using [For] Statement OPERATION INSTRUCTIONS Page IV .
Page 289: Com Communication Function
APPLIED OPERATIONS /Protocol processor 2-4-7 COM Communication function PAPP can communicate with RS-232C to the other PC. The data for sending and receiving should described with Macro commands. For detail of Macro command, see 2-4-2 [2 Table of commands]. Select COM port of PC. Click [Option] menu in [PAPP] window, then click [COM Control].
Page 290: Example Of Assembling A Protocol
APPLIED OPERATIONS / Protocol processor 2-4-8 Example of assembling a protocol This section describes an example of assembling a protocol. 1 Example of Setting Procedure of FRAP Observation An example of editing a protocol is described below assuming the FRAP observation. Protocol Repeat 30 times Image acquisition:...
Page 291 APPLIED OPERATIONS /Protocol processor Fig. 2-54 Panel After a Repeated Scan OPERATION INSTRUCTIONS Page IV . 2 - 1 7 7...
Page 292 APPLIED OPERATIONS / Protocol processor Displaying the Real-Time Graph (TIEMPO option software required) Specify the region in the image to display the real-time graph. For specifying method, refer to section 2-4-3, “Specifying the Regions Where Intensity Graph is Displayed in Real Time”. Display the [Tiempo] sub-panel in the [Acquire] panel to check the [Show live plot] check box.
Page 293 APPLIED OPERATIONS /Protocol processor Editing the Protocol Start and edit the protocol after setting of the observation condition has been completed. For starting method, refer to section 2-4-1, “Starting the Protocol Processor”. Fig. 2-56 Panel Displaying the [PAPP] window Set the [PAPP] window as shown below; Fig.
Page 294 APPLIED OPERATIONS / Protocol processor Set each Track as sown below; Setting Track 1 In XYT observation, set the zoom ratio to 2X and acquire an image before bleaching. Select [New mode]-“XYT” in [Mode] of [Track]. Select “Normal” in [Start by] of [Track]. Enter “1”...
Page 295 APPLIED OPERATIONS /Protocol processor Select “Series mode” in [Mode] of [Track]. Select “Normal” in [Start by] of [Track]. Enter “30” to [Condition] of [Track]. Enter “5” to [Frame Interval]. Enter “2” to [Zoom]. Select “Middle” in [Scan Speed]. (Return to the setting before bleaching.) Enter the laser intensity value for image acquisition to [Laser Intensity].
Page 296 APPLIED OPERATIONS / Protocol processor “ ” Protocol Repeat 30 times Get 5 Acquire Save Wait 5 Set Zoom Get one Wait 5 Acquire Set bleaching Set Zoom images Set Zoom image image power to x2 image bleach sec. sec. image portion power to x10...
Page 297 APPLIED OPERATIONS /Protocol processor Saving the Protocol Save the edited protocol data. The protocol can also be saved after being executed. Select [Save] or [Save as] in the [File] menu in the [PAPP] window. Save using the dialog box displayed. For details, see section 0, “...
Page 298 APPLIED OPERATIONS / Protocol processor Exiting the Protocol Terminate the protocol after acquiring the images. The images saved. Fig. 2-59 Panel When Exiting the Protocol OPERATION INSTRUCTIONS Page IV . 2 - 1 8 4...
Page 299: Example Of Setting Procedure Of Xyt Observation For Hours
APPLIED OPERATIONS /Protocol processor 2 Example of Setting Procedure of XYT Observation for hours (Protocol using repetition processing) An example of editing a protocol using the repetition processing is described below assuming the XYT observation including the GFP recovery observation for hours. Protocol 7 repeats Acquisition of...
Page 300 APPLIED OPERATIONS / Protocol processor One Point! The variable in the [For] statement can also be used by another cell in the protocol processor. <Setting example> It is also possible to change the laser intensity value at every image acquisition. In this case, assume that the laser intensity of the Ar laser is variable N and create a protocol for acquiring image as N varies from 10 to 100 in steps of 10.
Page 301 APPLIED OPERATIONS /Protocol processor Setting the Observation Condition Before setting and starting the XYT observation for hours using the Protocol Processor, execute a repeated scan and set the condition for the observation. Assuming FITC to Channel 2 and image size to 320X240 pixels, the following figure shows an example of image acquisition.
Page 302 APPLIED OPERATIONS / Protocol processor Editing the Protocol Start and edit the protocol after setting of the observation condition has been completed. For starting method, refer to section 2-4-1, “Starting the Protocol Processor”. Fig. 2-61 Panel Displaying the [PAPP] window Set the [PAPP] window as shown below;...
Page 303 APPLIED OPERATIONS /Protocol processor Set each Track as sown below; Setting Track 1 Acquire nine images in XYT observation. Select “New mode”-“XYT” in [Mode] of [Track]. Select “Normal” in [Start by] of [Track]. Enter “9” to [Condition] of [Track]. Select “Middle” in [Scan Speed]. Enter the laser intensity value for image acquisition to [Laser Intensity].
Page 304 APPLIED OPERATIONS / Protocol processor Enter the PMT value of each channel in the cell for each channel in [PMT Voltage]. Enter “Init” in [PMT2] and [PMT3]. Enter the image file name to be stored to [Save FileName.] Setting Track 4 Specifies the repeated protocol by use N variable.
Page 305 APPLIED OPERATIONS /Protocol processor One Point! XYZT observation protocol becomes available when “New mode”-“XYZT” is selected instead of selecting “New mode” – XYT on [Mode] of [Track]. Saving the Protocol Save the protocol edited. The protocol can also be saved after being executed. Select [Save] or [Save as] in the [File] menu in the [PAPP] window.
Page 306 APPLIED OPERATIONS / Protocol processor Exiting the Protocol After the initial image has been acquired and saved, “saving the image acquired in Track 4 in Track 5” is repeated for 7 times, after which the protocol terminates. The images saved Fig.
Page 307: Pop-Up Menu
APPLIED OPERATIONS /Protocol processor 2-4-9 Pop-up Menu Right-clicking the mouse in the cells in the [PAPP] window displays the pop-up menu to edit the protocol as sown below; Removes the effect of the previous operation. Repeats the previous operation. Copies the value in the selected cell. Cuts the value in the selected cell.
Page 308: Restrictions Of [Papp] Setting
APPLIED OPERATIONS / Protocol processor 2-4-10 Restrictions of [PAPP] setting 1 Restrictions with [Mode] and I/O card There are restrictions at Sub Mode and Angle setting in [PAPP] that comes from [Mode] and I/O card in use. The following table shows the restriction. Sub Mode Row Angle Row Mode...
Page 309 APPLIED OPERATIONS /Protocol processor XYZT Normal Clip ZoomIn Seq_Normal Seq_Clip Seq_ZoomIn LineSeq_Normal LineSeq_Clip LineSeq_ZoomIn Normal Clip ZoomIn FreeLine Fast FastClip LineSeq_Normal LineSeq_Clip LineSeq_ZoomIn Normal Clip ZoomIn FreeLine LineSeq_Normal LineSeq_Clip LineSeq_ZoomIn Normal Clip ZoomIn FreeLine LineSeq_Normal LineSeq_Clip LineSeq_ZoomIn Pt*T Normal O* Input is possible when combiner is FV5-COMBA only. OPERATION INSTRUCTIONS Page IV .
Page 310 APPLIED OPERATIONS / Protocol processor It is possible to check which I/O board is being used by the following method. In case that I/O2 is used, [Pan][Zoom] group boxes on [Acquire] panel cannot be selected because arrows are grayed out as shown below when the board is started.
Page 311: Restrictions With [Mode] And [Sub Mode]
APPLIED OPERATIONS /Protocol processor 2 Restrictions with [Mode] and [Sub Mode] There are restrictions at Mode and its Sub Mode setting in [PAPP]. The following table shows the restriction. Scan Status Row Mode Sub Mode X Pos Y Pos X Size Y Size Normal Clip...
Page 312 APPLIED OPERATIONS / Protocol processor Normal Clip ZoomIn FreeLine Fast FastClip LineSeq_Normal LineSeq_Clip LineSeq_ZoomIn Normal Clip ZoomIn FreeLine LineSeq_Normal LineSeq_Clip LineSeq_ZoomIn Normal Clip ZoomIn FreeLine LineSeq_Normal LineSeq_Clip LineSeq_ZoomIn Pt*T Normal O means can use the combination. X means can not set O* Input is possible when combiner is FV5-COMBA only.
Page 313: Changing The Image Display Method
APPLIED OPERATIONS /Changing the Image Display Method 2-5 Changing the Image Display Method The method of displaying an image acquired by observation or opened from a file can be changed as described below. 2-5-1 Displaying an Image in Simulated Colors Display the [Display] panel of the image to be colored at the front.
Page 314 APPLIED OPERATIONS /Changing the Image Display Method One Point! The [Color Tool] dialog box can also be displayed by a mouse operation. Display the image to be colored at the front of the [Display] panel, and right-click a point in the image. A pop-up menu as shown below is displayed.
Page 315: Editing The Lut (Look Up Table)
APPLIED OPERATIONS /Changing the Image Display Method 2-5-2 Editing the LUT (Look Up Table) An original LUT can be created by editing an existing LUT. 2-5-2-1 LUT Graph Editing According to Colors Display the [Display] panel of the image that you want to edit the LUT. Click the <LUT>...
Page 316 APPLIED OPERATIONS /Changing the Image Display Method Click the <Graph display> button at the top right of the [Color Tool] dialog box again. Finally, click the <OK> button to exit from the LUT editing. One Point! The [Color Tool] dialog box can also be displayed by a mouse operation. Display the image to be colored at the front of the [Display] panel, and right-click a point in the image.
Page 317: Lut Graph Editing By Gamma Correction
APPLIED OPERATIONS /Changing the Image Display Method 2-5-2-2 LUT Graph Editing by Gamma Correction The intensity data of an image can be reallocated to make it easier to view. Display the [Display] panel of the image to be subjected to LUT change. <LUT>...
Page 318: Switching The Displayed Channels (Ch1, Ch2, Ch3)
APPLIED OPERATIONS /Changing the Image Display Method To load a previously saved LUT, click <Load LUT>. 2-5-3 Switching the Displayed Channels (Ch1, Ch2, Ch3) The buttons on the bottom left of the screen can be used to select where the image of a single channel or images of multiple channels are to be displayed.
Page 319: Displaying Images Of Multiple Channels Simultaneously
APPLIED OPERATIONS /Changing the Image Display Method 2-5-4 Displaying Images of Multiple Channels Simultaneously (Side By Side VIews, Over And Under Views. Single View) Images from multiple channels can be displayed either by merging them or placing them side by side. It is also possible to display the image of only one of these channels. Use the buttons displayed at the top of the [Display] panel and those on the bottom right which are displayed when the corresponding image is clicked.
Page 320 APPLIED OPERATIONS /Changing the Image Display Method From the displayed list of buttons, click one of the <Side by side views> button or <Over And Under views> button. The icon shown in the <Display switching> button <Side by side views> buttons will change to the icons of the <Side by side views>...
Page 321: Displaying Merged Image Of Multiple Channels (Single Views)
APPLIED OPERATIONS /Changing the Image Display Method 2-5-4-2 Displaying Merged Image of Multiple Channels (Single Views) Display the [Display] panel of the merged image of multiple channels at the front. Click the <Display switching> button at the top of the [Display] panel. The list of <Display switching>...
Page 322: Changing The Number Of Divided Images
APPLIED OPERATIONS /Changing the Image Display Method 2-5-5 Changing the Number of Divided Images The number of images viewed simultaneously can be changed. Increased image is only to be displayed. The image increased in Add View is not subjected to these operations described below. 2-5-5-1 Increasing the Number of Divided Images 1.
Page 323 APPLIED OPERATIONS /Changing the Image Display Method 3. Select [Views], then select [Add a view] in the sub-menu. 4. A view is added to the rightmost position on the [Display] panel. Fig. 2-71 [Display] Panel After View Addition Up to 6 views can be displayed at once. OPERATION INSTRUCTIONS Page IV .
Page 324: Decreasing The Number Of Divided Images
APPLIED OPERATIONS /Changing the Image Display Method 2-5-5-2 Decreasing the Number of Divided Images Display the [Display] panel of the image to be changed at the front. Fig. 2-72 [Display] panel Right-click the image. A pop-up menu as shown below appears. Select [Views], then select [Remove a last view] in the sub-menu.
Page 325 APPLIED OPERATIONS /Changing the Image Display Method A view is removed from the [Display] panel. Fig 2-73 [Display] panel after View Removal OPERATION INSTRUCTIONS Page IV . 2 - 2 1 1...
Page 326: Switching The Display Method Of Multiple Images
APPLIED OPERATIONS /Changing the Image Display Method 2-5-6 Switching the Display Method of Multiple Images With images composed of multiple slices, such as time-lapse images or images acquired by changing the cross-sections, the image to be displayed at the front position can be switched or the images can be displayed successively.
Page 327: Displaying Multiple Image Slices Together
APPLIED OPERATIONS /Changing the Image Display Method Click and hold the <Display> button for successive display. To stop it, click the <Display> button again. Simply clock the <Display> button for frame-by-frame display. 2-5-7 Displaying Multiple Image Slices Together With images composed of multiple slices, such as time-lapse images or images acquired by changing the cross-sections, the image slices can be displayed together for simultaneous viewing.
Page 328: Displaying Multiple Images Per Channel
APPLIED OPERATIONS /Changing the Image Display Method 2-5-7-1 Displaying Multiple Images Per Channel Display the [Display] panel of one the images which are to be displayed together. The icon of the image is displayed in the frame at the top left of the [Tile] panel and the acquisition parameters used in image acquisition are displayed in the [Experiment] panel.
Page 329: Displaying Images Of Two Channels Together
APPLIED OPERATIONS /Changing the Image Display Method 2-5-7-2 Displaying Images of Two Channels Together Images acquired in a 2-channel mode can be displayed together for simultaneous view. The operation method is identical to the method for displaying images per channel except for the following point.
Page 330: Displaying Time-Lapse Images
APPLIED OPERATIONS /Changing the Image Display Method 2-5-7-3 Displaying Time-Lapse Images Multiple images acquired over time can be displayed side by side for simultaneous view. Display the [Display] panel of one of the time-lapse images to be displayed together. Set the number of images to be displayed together by using the < >...
Page 331: Displaying Multiple Cross-Section Images
APPLIED OPERATIONS /Changing the Image Display Method 2-5-7-4 Displaying Multiple Cross-Section Images Images acquired from different cross-sections can be displayed together for simultaneous view. The operation method is identical to the method for displaying time-lapse images except for the following point. See section 2-5-7-3, “Displaying Time-Lapse Images”. Select [Z] from the [Tile Over] drop-down list.
Page 332 APPLIED OPERATIONS /Changing the Image Display Method Click the <New Page> button. A new [Display] panel appears showing the same images as the displayed image. The number of the displayed images is as set in step 2 above. NOTE Use the <Retile> button when it is required to re-arrange the images in the currently displayed [Display] panel.
Page 333: Re-Arranging Images Using The Same Display Method
APPLIED OPERATIONS /Changing the Image Display Method 2-5-7-6 Re-arranging Images Using the Same Display Method All of the images displayed together in a [Display] panel can be rearranged simultaneously based on the same display method (channels, magnification, scroller position). Click one of the images displayed together. Two sets of buttons appear above and below the clicked image.
Page 334 APPLIED OPERATIONS /Changing the Image Display Method Click the <Experiment List> button in the toolbar at the bottom of the [Tile] panel. The [Experiments in Memory] dialog box appears as shown below. <Experiment List> button Fig. 2-80 [Experiments in Memory] Dialog Box From the [Experiments in Memory] dialog box, select the file name of the second image to be displayed and drag it into the frame at the top right of the [Tile] panel.
Page 335 APPLIED OPERATIONS /Changing the Image Display Method When displaying images acquired in a multi-channel mode, select the display method from the [Display Type] drop-down list. Click the <New Page> button. A new [Display] panel appears showing the two images one above the other. The image of the file displayed in the frame at the top left of the [Tile] panel is displayed on the upper part of the [Display] panel, and that of the file displayed in the frame at the top right is displayed on the lower part.
Page 336: Displaying An Image In Full Screen
APPLIED OPERATIONS /Changing the Image Display Method 2-5-8 Displaying an Image in Full Screen Only the image itself can be displayed to fill the screen by erasing all other display components such as the toolbar, panel and status bar. This feature is useful for taking pictures using an analog printer for creation of a slide.
Page 337 APPLIED OPERATIONS /Changing the Image Display Method One Point! The image can also be displayed full screen by the mouse operation on the image. Display the [Display] panel of the image to be displayed full screen, and click the mouse right button on the image. A pop-up menu as shown below appears.
Page 338: Magnifying/Reducing An Image
APPLIED OPERATIONS /Changing the Image Display Method 2-5-9 Magnifying/Reducing an Image The image can be magnified or reduced using the buttons displayed at the top of the [Display] panel. Magnification or reduction up to 3:1 or 1:3 the original image is possible. Display the [Display] panel of the image to be magnified or reduced.
Page 339: Image Processing
APPLIED OPERATIONS /Image Processing 2-6 Image Processing Images can be processed using the [Process] panel. Display the [Process] panel at the front. 2-6-1 Filtering Use the [Filters] sub-panel in the [Process] panel to apply filtering to images. Display the [Filters] sub-panel in the [Process] panel at the front. Icon of the image being displayed (Image to be processed by filtering).
Page 340: Contour Enhancement
APPLIED OPERATIONS /Image Processing 2-6-1-1 Contour Enhancement When an image is blurred by the boundaries between image grains becoming unclear, it can be sharpened by applying contour enhancement. Five types of filters are provided for use in the contour enhancement. 1 Laplacian filter This filter enhances the contours of the image grains.
Page 341 APPLIED OPERATIONS /Image Processing Fig. 2-85 Panel Displaying the Laplacian Filtered Image 2 Sobel filter This filter enhances the contours of the image grains. If the image contains noise, the noise is also enhanced. It has two filter formats, X and Y, as shown below. The format providing the larger value after filtering is used.
Page 342 APPLIED OPERATIONS /Image Processing 3 High-pass X filter The HIGH-PASS X filter passes the high-frequency structures in the X-direction of image. In this way, it can extract details by detecting positions with large variation. This processing is useful for making structures clear or extract the edges. The filter format is as shown below.
Page 343: Noise Reduction
APPLIED OPERATIONS /Image Processing 5 Prewitt filter This filter enhances the contours of image grains in a similar way to the Sobel filter, but more strongly than it. It has two filter formats, X and Y, as shown below. The format providing the larger value after filtering is used.
Page 344: Image Sharpening
APPLIED OPERATIONS /Image Processing 2 Median filter The Median filter reduces noise in image while leaving the edges intact. However, it may be ineffective in case noise is concentrated in some positions or the image is very noisy. The operation method is identical to Laplacian filtering except for the following point. See section 2-6-1-1-1, “Laplacian filter”.
Page 345: Correcting Dic Level Irregularities
APPLIED OPERATIONS /Image Processing 2-6-1-4 Correcting DIC Level Irregularities DIC level irregularities refers to uneven brightness of the image background which may be observed when a transmitted image is acquired in IDC observation. The DIC level irregularities can be corrected to make the image easier to view. If fluorescence observation is used, note that the correction is possible only with the transmitted images.
Page 346 APPLIED OPERATIONS /Image Processing Click the <DIC leveling> button. The [Level] panel is newly created in the [Display panel] and shows an image after the correction of the DIC level irregularities of the background. The background irregularities are solved. Fig. 2-87 [Level] Panel After DIC Level Correction If the DIC level irregularities cannot be reduced, repeat steps 1 to 3 above by varying the correction factor.
Page 347: Contrast Conversion
APPLIED OPERATIONS /Image Processing 2-6-2 Contrast Conversion The LUT intensity can be mapped (re-assigned) while observing a histogram. Mapping (re-assignment) results in changing the image contrast. An image acquired by observation contains intensity information in values from 0 to 4095, but the intensity information used in actual display takes values from 0 to 255 by assigning the original values from 0 to 4095 to values from 0 to 255 usually.
Page 348 APPLIED OPERATIONS /Image Processing When the image was acquired in the multi-channel mode, select whether the LUT <Display channel switch> intensity mapping (re-assignment) is applied to multiple channels simultaneously or buttons to a single channel. To select the target channel(s), use the <Display channel switch> buttons. The histogram of the selected channel(s) is displayed.
Page 349 APPLIED OPERATIONS /Image Processing Enclose the histogram section of interest using the scale in the [Master View] field. The magnified view of the region selected by the scale is shown in the [Selected View] field. vertical chart axis represents the degree and horizontal axis represents...
Page 350: Mathematical Operations Between Images
APPLIED OPERATIONS /Image Processing 2-6-3 Mathematical Operations Between Images Arithmetic or logical operations can be applied between two different images or between an image and a constant. 2-6-3-1 Image Addition Addition of an image to an image (constant to an image) is possible as described below. Display the [Math] sub-panel of the [Process] panel.
Page 351 APPLIED OPERATIONS /Image Processing Click the <Experiment List> button in the toolbar at the bottom of the [Process] panel. The [Experiments in Memory] dialog box appears as shown below. <Experiment List> button Fig. 2-92 [Experiments in Memory] Dialog Box From the [Experiments in Memory] dialog box, select the file name of the first image and drag it to the frame at the top left of the [Process] panel.
Page 352 APPLIED OPERATIONS /Image Processing Enter the constant for use in operation in the [Scalar Value] text box in the [Scalar Operations] group box. (This step is required only for operation between an image and a constant.) To add an image to an image: Click the <Add 2 Images>...
Page 353: Image Subtract
APPLIED OPERATIONS /Image Processing 2-6-3-2 Image Subtract Subtraction of an image from an image (constant from an image) is possible as described below. The operation method is identical to Image + Image (Image + Constant) except for the following point. See section 2-6-3-1, “Image + Image (Image + Constant)”. To subtract an image from an image: Click the <Subtract 2 Images>...
Page 354: Not Image
APPLIED OPERATIONS /Image Processing To divide an image by an image: Click the <Divide 2 Images> button in the [Multi Image Operations] group box. A <Divide 2 Images> new [Display] panel showing [Image/Image] in the page tab appears, displaying the button image obtained by the division operation.
Page 355 APPLIED OPERATIONS /Image Processing Click the <NOT Image> button. A new [Display] panel showing [NOT] in the page <NOT Image> tab appears, showing the image obtained by the operation. button Fig. 2-95 [NOT] Panel OPERATION INSTRUCTIONS Page IV . 2 - 2 4 1...
Page 356: Image And Image
APPLIED OPERATIONS /Image Processing 2-6-3-6 Image AND Image Two different images can be ANDed. Display the [Math] sub-panel of the [Process] panel at the front. Sets the image to be subjected to Sets the second image operation. The icon of the image of the operation.
Page 357 APPLIED OPERATIONS /Image Processing From the [Experiments in Memory] dialog box, select the file name of the first image and drag it to the frame at the top left of the [Process] panel. The icon of the image is displayed in the frame at the top left of the [Process] panel. Before the dragging and dropping, the frame at the top left shows the icon of the image file displayed in the [Display] panel.
Page 358: Image Or Image
APPLIED OPERATIONS /Image Processing Click the <Image AND Image> button. A new [Display] panel showing [Image AND <Image AND Image> Image] in the page tab appears, showing the image obtained by the operation. button Fig. 2-98 [Image AND Image] Panel 2-6-3-7 Image OR Image Two different images can be ORed.
Page 359: Brightness Overlap Level Between 2 Channels (Colocalization)
APPLIED OPERATIONS /Image Processing 2-6-4 Brightness overlap level between 2 channels (Colocalization) This function enables to observe overlap intensity of two channel of image. Acquires or opens images which more than two channels, and displays the images on [Display] panel. On the [Display] panel, displays and overlaps 2 channel image that to be observed.
Page 360: Annotation Mode
APPLIED OPERATIONS /Image Processing 2-6-4-1 Annotation Mode 1 None Mode None Mode is the default of [Colocalization Processor] dialog box. The mode can be changed by option button in [Annotation Mode] Group Box. The items described for this mode can be used in other modes. [Data Selection] Group Box <Disabled>...
Page 361 APPLIED OPERATIONS /Image Processing 2 Thresholds Mode Specifies Threshold level on [Histogram] graph, then measures by use of the threshold level. [Histogram] Graph Threshold line vertical horizontal red line) appears. The value can be adjust by dragging those lines. [Annotation mode] Group [Annotation] Text Box When <Thresholds>...
Page 362 APPLIED OPERATIONS /Image Processing [Histogram] graph Explanation of [Annotation] Text Box Display in Text Box Explanation Threshold Annotation Threshold Mode Total samples in histogram: Total numbers of image pixels to be processed (Dye-name1)[Thresh: Name of Dye method. Threshold value in X- axis.
Page 363 APPLIED OPERATIONS /Image Processing 3 Min-Max Bounds Mode Specifies the rectangular on [Histogram] graph, and measures colocalization by use of the rectangular. In addition, distribution level of brightness inside rectangular is displayed on [Display] panel. Color is the same as color of rectangular selected. [Histogram] Graph Rectangle is displayed in graph.
Page 364 APPLIED OPERATIONS /Image Processing Explanation of [Annotation] Text Box Display in Text Box Explanation Min-Max Annotation Min-Max Bounds Mode Total samples in histogram Total numbers of image pixels to be processed (Dye-name1) limits: X-axis range of rectangular Minimum: Minimum value of X-axis rectangular Maximum: Maximum value of X-axis rectangular (Dye-name2) limits:...
Page 365 APPLIED OPERATIONS /Image Processing 4 Regions Mode Specifies the arbitrary region in [Histogram], and measures colocalization within the region. In addition, distribution level of brightness in draw pictorial figure is displayed on [Display] panel. Color is the same as color of draw pictorial figure selected. [Histogram] Graph You can draw a pictorial figure on [Histogram], using graphic...
Page 366 APPLIED OPERATIONS /Image Processing Explanation of [Annotation] Text Box (In case that 3 pictorial figures exist. Items (1) will be increased according to number of pictorial figures.). Display in Text Box Explanation Regions Annotation Regions Mode Total samples in histogram Total numbers of image pixels to be processed RGB:...
Page 367: Colocalization For Series Image Data Set
APPLIED OPERATIONS /Image Processing 2-6-4-2 Colocalization for series image data set Colocalization is also available for series image data set, i.e XYZ image data set. [Data Selection] Group Box <Current slice> Option Button The cocalization is processed for the image in a series data set which is displayed in [Display] panel.
Page 368 APPLIED OPERATIONS /Image Processing 2-6-4-3 Image measurement When option button of [Image Measures] of [Analysis Mode] Group Box is selected, the dialog as shown below appears. [Analysis Mode] Group Box [Image Measurements] Group Box It displays results of measurement in text box below.
Page 369 APPLIED OPERATIONS /Image Processing ・Pearson’s Coefficient： In case that the value to be acquired is defined as However, legends used for image measurement are as follows: : Brightness of wavelength -th pixel, : Brightness of wavelength -th pixel : Average brightness of wavelength : Average brightness of wavelength ・Overlap：...
Page 370 APPLIED OPERATIONS /Image Processing However, should be -th pixel that belongs to aggregation A that is greater than threhold value. OPERATION INSTRUCTIONS IV . Page 2 - 2 5 6...
Page 371: Appending Image (Append)
APPLIED OPERATIONS /Image Processing 2-6-5 Appending image (Append) 2-6-5-1 Appending two images Two images (A and B) acquired in XY or XYZ observation can be appended along the Z- or T-direction or as animation. Examples) XY image Appending in XYT image T-direction TIME XY image...
Page 372 APPLIED OPERATIONS /Image Processing Two XY images can be appended in the T-direction with the following procedure. Open the two image files (A and B) for appending. Display the [display] panel of the first image (A) at the front. Display the [Experiment Editor] sub-panel of the [Process] panel at the front. Fig.
Page 373 APPLIED OPERATIONS /Image Processing In the toolbar at the bottom of the [Process] panel, click the <Experiment List> button. The [Experiments in Memory] dialog box appears as shown below. <Experiment List> button Fig. 2-106 [Experiments in Memory] Dialog Box The icon of the image file being displayed at the front of the [display] panel (A) is shown in the frame at the top left of the [Process] panel.
Page 374 APPLIED OPERATIONS /Image Processing In the [Experiments in Memory] dialog box, select the file name of the second image (B) and drag it to the frame at the top right of the [Process] panel. The icon of the second image will be displayed in the frame at the top right of the [Process] panel.
Page 375 APPLIED OPERATIONS /Image Processing A new [display] panel having [Append] as the window title is created, showing the image after appending. For appending more than 3 images at a time, see section 2-6-5-2. OPERATION INSTRUCTIONS Page IV . 2 - 2 6 1...
Page 376: Appending Image From Several Image Data Set
APPLIED OPERATIONS /Image Processing 2-6-5-2 Appending image from several image data set An image data set can be made from appending several image data set. NOTE There are the following restrictions when you append image files. ·Two image files must be the same image size. ·Two image files must be the same channel number.
Page 377 APPLIED OPERATIONS /Image Processing [Append on New dimension] dialog box appears as shown below. Image Info area File area [Order] Text Box Folder area <Clear> Button <Append> Button <Brows> Button [Created file] [Resolution] [Dimension] Text Box Text Box Group Box [Comments] [Sort] Text Box...
Page 378 APPLIED OPERATIONS /Image Processing Select Series data set type that will be created in [Dimension] Group Box. The option buttons work as the followings. ·on T ････････････ Appends file in T direction. ·on Z ････････････ Appends file in Z direction. ·on An ･･･････････...
Page 379 APPLIED OPERATIONS /Image Processing Enter folder and file name into [Create file] text box. By pressing <Browse…> button, folder appears for easy folder selection. Click <Append> button, then new appended image created and stored. Pressing <Clear> button clears the file assignment. [Resolution] text box works as follows.
Page 380 APPLIED OPERATIONS /Image Processing One Point! Plural numbers of images can be appended without starting FLUOVIEW software. 1. Select <Start> button at bottom of Windows screen. 2. In case of Windows NT, select [Programs] – [Windows NT Explorer] command from the [Start] menu.
Page 381: Extract Image (Crop)
APPLIED OPERATIONS /Image Processing 2-6-6 Extract image (Crop) 2-6-6-1 Extract image data from an image data set New image data with selected slice image from an image data set can be made by this function. The following is an example for Image extraction from an XYZT image data set. 1.
Page 382 APPLIED OPERATIONS /Image Processing The following method of slice selection is also available. ·All of image slices can be select by pressing <select all> button. ·All of row or column slices can be selected by pressing [No. Display]. ·Image slices with a certain time interval and/or Z step can be selected by pressing <Set>...
Page 383 APPLIED OPERATIONS /Image Processing One Point! When arbitrary portion of XYT image is extracted and plural numbers of [No. Display] columns at T columns are selected, the dialog box as shown below will appear in case that number of units at Z portion of [No. Display] on all T columns are not the same number of units.
Page 384 APPLIED OPERATIONS /Image Processing One Point! It is possible to cut out the image and extract it by specifying arbitrary region. Image cut out is done as follows. <Annotate> button 1. Select [Annotate] button in tool bar. ＜Circle＞ button ＜Rectangular＞ button ＜Free Region＞...
Page 385 APPLIED OPERATIONS /Image Processing One Point! Slice extract function works even if FLUOVIEW software is not running. However, clipping function works only with FLUOVIEW software. 1.Press Windows <Start> button. 2.In case of Windows NT, select [Windows NT Explorer] command in [Programs]. In case of Windows 2000, select [Windows Explorer] command in [Accessories].
Page 387: Image Analysis
APPLIED OPERATIONS /Image Analysis 2-7 Image Analysis Images can be analyzed using the [Analyze] panel. Display the [Analyze] panel at the front. Displays the icon of the image being displayed (image to be subjected to <Begin Analysis> button analysis). Starts analysis. [Measurement Results] box Shows the measurement data of the specified line or region.
Page 388: Checking The Intensity Of A Specific Part
APPLIED OPERATIONS /Image Analysis 2-7-1 Checking the Intensity of a Specific Part 2-7-1-1 Intensity Values on a Line (Line Profile) The intensity values on a line in an image can be displayed graphically. Display the [Single] sub-panel at the front. Display the [Display] panel of the image to be subjected to the intensity checking at the front.
Page 389 APPLIED OPERATIONS /Image Analysis To specify a free line: On the image, drag the mouse pointer along the line <Free Line> button to be checked. The line is displayed on the image together with the handles on it. The intensity profile can be displayed Handle while the handles are displayed.
Page 390 APPLIED OPERATIONS /Image Analysis Double-click the [Intensity Profile] button. The [Enhanced Profile] window appears as shown below. <Properties> buton Displays the [Editing] dialog box for use in detailed setting of the chart or change of the chart display. See section 2-15, “Changing the Chart Display Method”...
Page 391: Intensity Values On A Planar Region (Bird's Eye View)
APPLIED OPERATIONS /Image Analysis 2-7-1-2 Intensity Values on a Planar Region (Bird’s Eye View) The intensity values on a region in an image can be displayed graphically. Display the [Single] sub-panel at the front. Display the [Display] panel of the image to be subjected to the intensity checking at the front.
Page 392 APPLIED OPERATIONS /Image Analysis Specify the region to be checked in the image in the [Display] panel. They can be specified as described below. To specify a rectangle: On the image, drag the mouse pointer along the diagonal line of the desired rectangle, from the top left corner to the bottom right corner.
Page 393 APPLIED OPERATIONS /Image Analysis Click the <Annotate> button so that the list of buttons disappears. Click the <Begin Analysis> button. The bird’s eye view of the specified region will <Annotate> button be displayed in the [Intensity Profile] box of the [Analyze] panel. The [Measurement Results] box in the [Single] sub-panel shows the measurement results such as the area (Area), horizontal and vertical lengths ( X/Y, Z or T) and perimeter length (Perimeter) of the region,...
Page 394 APPLIED OPERATIONS /Image Analysis 10. Double-click the [Intensity Profile] button. The [Intensity Map] window appears as shown below. [Angle] scale Sets the angle in the horizontal direction. The result can be confirmed with the small bird’s eye view in the frame on the [Tilt] scale Sets the angle in the vertical direction.
Page 395: Checking The Intensity Distribution Of A Specific Part
APPLIED OPERATIONS /Image Analysis 2-7-2 Checking the Intensity Distribution of a Specific Part 2-7-2-1 Intensity Distribution on a Line (Histogram) The histogram on a line in an image can be displayed. The histogram is displayed in the [Region Histogram] box in the [Single] sub-panel. The operation method is identical to displaying the intensity profile on a line.
Page 396: Intensity Distribution On A Planar Region (Histogram)
APPLIED OPERATIONS /Image Analysis The magnification or scrolling of the graph can be canceled by dragging the left button of the mouse from the bottom left to the right of the magnified graph. When the mouse pointer is placed on a graph line while the Ctrl Alt key is held depressed, the coordinates can be displayed.
Page 397 APPLIED OPERATIONS /Image Analysis When a desired area is specified by dragging the left button of the mouse on the graph, the specified area can be magnified. When the right button of the mouse is dragged on the graph, the graph can be scrolled.
Page 398: Measuring A Part
APPLIED OPERATIONS /Image Analysis 2-7-3 Measuring a Part 2-7-3-1 Length Measurement The length between two points in an image or the perimeter of a region in an image can be measured. The [Measurement Results] box in the [Single] sub-panel shows the measurement results such as the length between 2 points (Length) or perimeter of the region (Perimeter) and the horizontal and vertical lengths (X/Y, Z or T) of the region, and the statistic result of each channel such as the total (Total), average (Average) and standard...
Page 399: Measuring The Change In Mean Value Of Intensity
APPLIED OPERATIONS /Image Analysis 2-7-3-3 Measuring the Change in Mean Value of Intensity The mean value of the intensity in a region specified in an image can be measured and displayed graphically. Display the [Series] sub-panel at the front. Display the [Display] panel of the image that you want to check the intensity at the front.
Page 400 APPLIED OPERATIONS /Image Analysis hen the image is composed of multiple image slices, the range of image slices to be subjected to the operation can be set using the <Set start position> button <Set start position> and <set end position> buttons above the images. First display the image slice to start the operation using the <Display>...
Page 401 APPLIED OPERATIONS /Image Analysis W Click the <Annotate> button in the toolbar at the bottom of the [Analyze] panel. A list of buttons appear as shown below. <Annotate> button <Rectangular> button <Circle> button < Polyregion > button < Free Region> button From the displayed buttons, click the <Rectangular>...
Page 402 APPLIED OPERATIONS /Image Analysis Click the mouse. A pop-up menu as shown below appears. Select [Properties] from the menu. The [Properties] dialog box as shown below appears. Display the [Color] panel at the front. Fig 2-116 [Properties] Dialog Box 10. Select the desired color from the [Color Palette] list box. 11.
Page 403 APPLIED OPERATIONS /Image Analysis 14. Click the <Begin Analysis> button. The mean value of the specified regions will be displayed graphically in the [Mean Intensity] box. NOTE The colors of the chart lines corresponding to the colors assigned to the regions. NOTE When the image was acquired in the multi-channel mode, the channel number is displayed to the right of each chart line.
Page 404 APPLIED OPERATIONS /Image Analysis 15. Double-click the [Mean Intensity] box. The [Average Intensity Trace] window appears as shown below. <Properties> button Displays the [Editing] dialog box for use in detailed setting of the chart or change of the chart display. See section 2-15, “Changing the Chart Display Method”...
Page 405: Measuring The Change In Integrated Intensity
APPLIED OPERATIONS /Image Analysis 2-7-3-4 Measuring the Change in Integrated Intensity The total value of the intensity in a region specified in an image can be measured and displayed graphically. The operation results are displayed graphically in the [Integrated Intensity] box in the [Series] sub-panel.
Page 406 APPLIED OPERATIONS /Image Analysis <Properties> button Displays the [Editing] dialog box for use in detailed setting of the chart or change of the chart display. See section 2-15, “Changing the Chart Display Method” for details. <Copy> button Copies the plotted image in the clipboard. <Save>...
Page 407: Building An Image From A Different Viewpoint
APPLIED OPERATIONS /Building an Image from a Different Viewpoint 2-8 Building an Image from a Different Viewpoint 2-8-1 Building Extended Focus Image from XYZ Image 2-8-1-1 Display Switching to Built Image An extended focus image can be built from XYZ (multiple sections) images and the display can be switched to show the built image.
Page 408 APPLIED OPERATIONS /Building an Image from a Different Viewpoint (Image No. 0) (Image No. 1) (Image No.2) (Image No. 3) Fig. 2-120 Four Images Used in Building Extended-Focus Image Fig. 2-121 Panel Showing an Extended-Focus Image OPERATION INSTRUCTIONS IV . Page 2 - 2 9 4...
Page 409 APPLIED OPERATIONS /Building an Image from a Different Viewpoint Among the multiple image slices composing the XYZ image, the range of image slices to be used in extend image building can be specified. Display the image slice to be set as the start image by using the <Display> buttons at the top of the [Display] panel.
Page 410: Turning Built Image Into Single Image
APPLIED OPERATIONS /Building an Image from a Different Viewpoint 2-8-1-2 Turning Built Image into Single Image From XYZ (multiple sections) image, an extended-focus image can be built as a separate image from the original image. Use the [Visualize] panel to build the image. First display the [Visualize] panel.
Page 411 APPLIED OPERATIONS /Building an Image from a Different Viewpoint Fig. 2-123 Panel Showing Extended-Focus Image OPERATION INSTRUCTIONS Page IV . 2 - 2 9 7...
Page 412: Turning Built Image Into Time Series Image
APPLIED OPERATIONS /Building an Image from a Different Viewpoint 2-8-1-3 Turning Built Image into time series image From XYZT image, an extended-focus image can be built as a separate time series image from the original image. Use the [Visualize] panel to build the image. First display the [Visualize] panel.
Page 413 APPLIED OPERATIONS /Building an Image from a Different Viewpoint Display the [Display] panel of the XYZ (multiple sections) image. Click the <Make Multi Plane View> option button in the [Rendered View] panel. <Make Multi Plane View> The [3D-] panel is created to start the line image building. button Line image display in XZ-direction Display the image of the line...
Page 414: Viewing 3D Image
APPLIED OPERATIONS /Viewing 3D Image 2-9 Viewing 3D Image Use the [Visualize] panel to view an image three-dimensionally. First display the [Visualize] panel. [Display] panel Shows the image. The file name of the [Begin Visualize] button image is shown in the page tab of the Starts building the images for 3D panel.
Page 415 APPLIED OPERATIONS /Viewing 3D Image Fig 2-129 Example of Intensity Projection Fig 2-130 Example of Topographic Projection [Threshold range Low / High] scale Sets the range of intensity values to be used in image building. [Depth Weight] scale Sets the weighting in the depth direction. By increasing this setting, it is possible to provide the image with a perspective by darkening the far objects and brightening near objects.
Page 416: Successive Display Of Images
APPLIED OPERATIONS /Viewing 3D Image 2-9-1 Successive Display of Images Images composed of multiple image slices acquired by varying the multiple sections (XYZ observation, XYZT observation) can be displayed successively using the buttons at the top of the [Display] panel as shown below. Display the [Display] panel of the image composed of multiple image slices.
Page 417: Changing The Successive Display Speed
APPLIED OPERATIONS /Viewing 3D Image Click the <Set start position> button. (If the start position is not set, image No. 0 becomes the start image automatically.) Display the image slice to end the successive display by using the <Display> button at the top of the [Display] panel.
Page 418: Changing The Successive Image Display Position
APPLIED OPERATIONS /Viewing 3D Image Select the option button of the speed to be varied. Set the desired display speed in the scale on the right. Click the <OK> button to close the [Animation speed] dialog box. 2-9-1-2 Changing the successive image display position The successive display position of multiple image slices in an image can be changed using a bottom displayed at the top of the [display] panel.
Page 419 APPLIED OPERATIONS /Viewing 3D Image For the switching of channels, see section 2-5-3, “Switching the Display Channels”. Click the [Arbitrary View] option button in the [Rendered View] group box. Click the [Brightest] or [Sum] option button in the [Method] group box. Select the image rotation direction from the [Standard Views] drop-down list.
Page 420 APPLIED OPERATIONS /Viewing 3D Image During the building, the <Begin Visualize> button turns into the <Cancel <Cancel Visualize> Visualize> button. Click the <Cancel Visualize> button if you want to cancel button the building. Fig. 2-132 Panel Showing the Built Image 12.
Page 421: Building Stereo 3D Images
APPLIED OPERATIONS /Viewing 3D Image 2-9-3 Building Stereo 3D Images Images composed of multiple image slices acquired by varying the multiple sections (XYZ observation) can be built into a pair of stereo 3D images. These images can be viewed three-dimensionally by watching them with two eyes for a while. The operation is similar with animation.
Page 422 APPLIED OPERATIONS /Viewing 3D Image Fig. 2-133 Panel Showing Stereo 3D Images OPERATION INSTRUCTIONS IV . Page 2 - 3 0 8...
Page 423: Building A 3D Image To Be Viewed Through Color (Red/Green) Eyeglasses2-293
APPLIED OPERATIONS /Viewing 3D Image 2-9-4 Building a 3D Image to be Viewed Through Color (Red/Green) Eyeglasses Images composed of multiple image slices acquired by varying the multiple sections (XYZ observation) can be built into an image which looks 3D when viewed through a pair of color (red/green) eyeglasses.
Page 424 APPLIED OPERATIONS /Viewing 3D Image Fig. 2-134 Panel Showing 3D Image to be Viewed Through Color Eyeglasses OPERATION INSTRUCTIONS IV . Page 2 - 3 1 0...
Page 425: Viewing Images Following The Progress Of Time
APPLIED OPERATIONS /Viewing Images Following the Progress of Time 2-10 Viewing Images Following the Progress of Time Images composed of multiple image slices (XYT, XYZT or XZT observation) can be displayed following the time lapse to show the change over time. 2-10-1 Displaying Images Together The change over time can be viewed at a glance by displaying multiple image slices together.
Page 426 APPLIED OPERATIONS /Viewing Images Following the Progress of Time <Loop> button <Z/T series switch> button Successive display or frame-by-frame display is <Set start position> button repeated in the same direction. When successive display or frame-by-frame display is <Loop/Bound switch> buttons required, set the image with which the display should start.
Page 427: Transferring Data To Another Application
<Close> button Quits the [Enhanced Profile Plot] window and returns to the [Analyze] panel. Fig. 2-135 [Enhanced Profile Plot] Window Microsoft Excel is not included in the FLUOVIEW FV300 system. Please purchase it separately. OPERATION INSTRUCTIONS Page IV . 2 - 3 1 3...
Page 428 APPLIED OPERATIONS /Transferring Data to Another Application Click the <Save> button. When the [Save As] dialog box appears as shown below, set the file name and click the <OK> button to save the analysis data. Fig. 2-136 [Save As] Dialog Box Exit from FLUOVIEW or display the [Start] menu by pressing the Ctrl + keys.
Page 429 APPLIED OPERATIONS /Transferring Data to Another Application Click the <Next> button. When the dialog as shown below appears, check the [Tab] check box in the [Delimiters] group box, then select [[none]] from the [Text Qualifier:] drop-down list. Fig. 2-138 Dialog Box When the File is Opened by Excel 2 / 3 Click the <Next>...
Page 430: Transferring The Plot Image Of Analysis Data To Another Application
APPLIED OPERATIONS /Transferring Data to Another Application For detailed operation procedures of Excel, refer to the [Excel manuals]. 2-11-2 Transferring the Plot Image of Analysis Data to Another Application The plot image of analysis data can be transferred to an application handling images, such as Paint.
Page 431: Transferring Image Data To Another Application (Microvoxel, Paint, Etc.)2-301
APPLIED OPERATIONS /Transferring Data to Another Application Click the <Copy> button to copy the plot image to the clipboard. Exit from FLUOVIEW or display the [Start] menu by pressing the Windows key or Ctrl + Esc keys. Select [Programs]-[Accessories] and issue the [Paint] command. From the [Edit] menu of Paint, select the [Paste] command and paste the plot image which has been copied to the clipboard in step 3.
Page 432: Entering Comment In Image
APPLIED OPERATIONS /Entering Comment in Image 2-12 Entering Comment in Image Comment can be entered in an image for use in presentation or slide creation. Use the <Annotate> button in the toolbar at the bottom left of the screen. Click the <Annotate>...
Page 433 APPLIED OPERATIONS /Entering Comment in Image From the drop-down list in the dialog box, select one of the following labels. Z Position <z> Time Instant <t> Channel # <channel> Display Zoom <display zoom> Is bounded <bounds> Experiment Name <name> <display zoom> <z> <t> <animation> <channel> <stereo> <bounds> Desired characters can also be entered.
Page 434: Displaying The Image Intensity
APPLIED OPERATIONS /Entering Comment in Image Select the character font and size using the [Font], [Font Style] and [Size] list boxes. Click the <OK> button to close the [Font] dialog box. Place and click the mouse pointer on the image position you want to enter characters.
Page 435: Displaying The X-Coordinate/Y-Coordinate Of The Image
APPLIED OPERATIONS /Entering Comment in Image 2-12-3 Displaying the X-coordinate/Y-coordinate of the Image The X-coordinate position or the Y-coordinate position of any pixel of an image can be displayed. Display the [Display] panel of the image that you want to display the X-coordinate position or the Y-coordinate position at the front.
Page 436: Drawing A Figure In Image
APPLIED OPERATIONS /Entering Comment in Image 2-12-4 Drawing a Figure in Image This facility is used to draw figures in the image. FLUOVIEW provides seven figure drawing modes. Display the [Display] panel of the image in which you want to draw figures. Select one of the following command buttons and draw a figure in the image using the mouse.
Page 437: Drawing A Scale In Image
APPLIED OPERATIONS /Entering Comment in Image 2-12-5 Drawing a Scale in Image A scale can be drawn between two points in an image. Display the [Display] panel of the image in which you want to draw a scale. <Scale> button Click the <Scale>...
Page 438 APPLIED OPERATIONS /Entering Comment in Image The size of characters in the scale can also be changed. The size of characters in the scale is determined by the size of characters written using the <Text> button. Perform the following operation before starting to draw the scale.
Page 439: Drawing An Arrow In Image
APPLIED OPERATIONS /Entering Comment in Image 2-12-6 Drawing an Arrow in Image This facility is used to draw an arrow for indicating a point in interest in image or adding explanation in it. Display the [Display] panel of the image you want to draw an arrow. Click the <Arrow>...
Page 440: Drawing Color Bars In Image
APPLIED OPERATIONS /Entering Comment in Image 2-12-7 Drawing Color Bars in Image This facility is used to draw color bars in an image. Display the [Display] panel of the image you want to draw color bars. Click the <Color Bar> button in the displayed list of buttons. <Color Bar>...
Page 441: Deleting Comment
APPLIED OPERATIONS /Entering Comment in Image 2-12-8 Deleting Comment Click the mouse on the comment to be deleted to make the comment active (i.e. handles displayed around the comment). Handle Click the right button of the mouse. A pop-up menu as shown below appears. Fig.
Page 442: Changing The Comment Size
APPLIED OPERATIONS /Entering Comment in Image A comment can also be moved by selecting it, placing the mouse pointer on it so that the mouse pointer turns into a cross (+), then dragging the mouse. In this case, the final positioning of the comment can be determined by placing the mouse pointer outside the areas enclosed by the handles and clicking the left button of the mouse.
Page 443: Changing The Comment Color
APPLIED OPERATIONS /Entering Comment in Image 2-12-11 Changing the Comment Color Click the mouse on the comment to be changed of color to make the comment active (i.e. handles displayed around it). Click the right button of the mouse. A pop-up menu as shown in Fig. 2-145 appears. Select [Properties] from the menu.
Page 444: Changing The Comment Font
APPLIED OPERATIONS /Entering Comment in Image 2-12-12 Changing the Comment Font Click the mouse on the comment to be changed of font to make the comment active (i.e. handles displayed around it). Click the right button of the mouse. A pop-up menu as shown in Fig. 2-145 appears. Select [Properties] from the menu.
Page 445: Image Output At Printer
APPLIED OPERATIONS /Image Output at Printer 2-13 Image Output at Printer Display the [Display] panel of the image to be output at the printer. Click the <Print> button in the toolbar at the bottom left of the panel. A dialog box as shown below appears.
Page 446 APPLIED OPERATIONS /Image Output at Printer One Point! The [Print] dialog box can also be displayed by mouse operation on the image. Display the image to be output at the printer at the front of the [Display] panel, and click the right button of the mouse on the image. A pop-up menu as shown below appears.
Page 447: Merger/Extraction Of Image Channels
APPLIED OPERATIONS / Merger/Extraction of Image Channels 2-14 Merger/Extraction of Image Channels 2-14-1 Setting the Range of Multiple Image Slices When merging or extracting channels in images composed of multiple image slices, such as time-lapse images and images acquired by varying the cross-sections, it is possible to select only some of image slices as the target of channel merger or extraction.
Page 448: Merging Image Channels
APPLIED OPERATIONS /Merger/Extraction of Image Channels Display the image slice to start the range at the front using the <Display> button at the top of the [Display] panel. Click the <Set start position> button. (If the start position is not set, image No. 0 becomes the start image automatically.) Display the image slice to end the range at the front using the <Display>...
Page 449 APPLIED OPERATIONS /Merger/Extraction of Image Channels When the image in one of the image files is composed of multiple image slices, it is possible to use only some of the slices by setting a slice range. See section 2-14-1, “Setting the Range of Multiple Image Slices” for the operation procedure.
Page 450 APPLIED OPERATIONS /Merger/Extraction of Image Channels Click the <Merge channels> button. The [Experiments in Memory] dialog box appears as shown below. <Merge channels> button Fig. 2-150 [Experiment Editor] Group Box The frame at the top left of the [Experiments in Memory] dialog box shows the icon of the image file displayed at the front of the [Display] panel.
Page 451 APPLIED OPERATIONS /Merger/Extraction of Image Channels NOTE When the image of the second selected image file is composed of multiple image slices, the setting of the image slice range is ineffective even when the range is set. Up to 3 channels are selected automatically beginning with the first image set in the [Experiments in Memory] dialog box, and connected to [Merger] by lines.
Page 452 APPLIED OPERATIONS /Merger/Extraction of Image Channels Click the <Merge> button in the [Experiments in Memory] dialog box. A new [Display] panel showing [Merge] in the page tab appears and the images including newly merged channel(s) are displayed in the panel. Fig.
Page 453: Extracting Channels From Image
APPLIED OPERATIONS /Merger/Extraction of Image Channels 2-14-3 Extracting Channels from Image Desired channels can be extracted from an image. Use this facility to extract only the images of required channels from an image acquired from more than one channel. Open the image file of the image to extract channels in advance. When the image has been saved in more than one image file, it is possible to use only some of the slices by setting a slice range.
Page 454 APPLIED OPERATIONS /Merger/Extraction of Image Channels Click the <Extract channels> button. The [Experiments in Memory] dialog box appears as shown below. <Extract channels> button Fig. 2-153 [Experiment Editor] Group Box The frame at the top left of the [Experiments in Memory] dialog box shows the icon of the image file displayed at the front of the [Display] panel.
Page 455 APPLIED OPERATIONS /Merger/Extraction of Image Channels From the file list in the [Experiments in Memory] dialog box, select the file name of the image from which to extract channels and drag it into the frame at the top left. The icon of the image is displayed in the frame at the top left. The mouse pointer turns into the image icon during dragging.
Page 456 APPLIED OPERATIONS /Merger/Extraction of Image Channels Among the channel lines connected to [Extract], deselect the unnecessary channels as described in the TIP on the previous page so that only the necessary channels are selected. Click the <Extract> button in the [Experiments in Memory] dialog box. A new [Display] panel showing [Extract] in the page tab appears and the image of the extracted channel(s) is displayed in the panel.
Page 457: Changing The Chart Display Method
APPLIED OPERATIONS /Changing the Chart Display Method 2-15 Changing the Chart Display Method When the processed analysis data chart in the [Analyze] panel is double-clicked, the [Enhanced Profile Plot] dialog box appears. By clicking the <Properties> button, the [Editing] dialog box can be displayed, allowing the detailed chart settings and chart display method to be changed.
Page 458 APPLIED OPERATIONS /Changing the Chart Display Method [TeeChart Gallery] sub-panel Displayed when the <Add> or <Change> button is clicked. Used to set the chart type. The chart types that cannot be set cannot be selected. Enable or disable the 3D display.
Page 459 APPLIED OPERATIONS /Changing the Chart Display Method [Axis] sub-panel Used to set the coordinate axes of the chart. Enable or disables the display of chart axes. Display the chart axes by selecting the optimum scale automatically. maximum Select the chart axes to be set minimum values of the chart in the [Scales], [Title], [Labels], axes either automatically or...
Page 460 APPLIED OPERATIONS /Changing the Chart Display Method [Labels] sub-panel Used to set the label of the coordinate axis. Enable or disable the label display on a point of crossing with another axis. Enable or disable the label display. Set the label character font. Not used with the present Set the distance from the title application.
Page 461 APPLIED OPERATIONS /Changing the Chart Display Method [Position] sub-panel Used to set the coordinate axis display position. Set the coordinate axis display position with respect to the chart. Set the coordinate axis display range with reference to the start point. Set the coordinate axis display range...
Page 462 APPLIED OPERATIONS /Changing the Chart Display Method [Legend] sub-panel Used to set the chart legend display. Set the items to be Enable or disable the legend displayed in the legend. Usually set [Automatic]. display. Set the legend display Set the legend background style.
Page 463 APPLIED OPERATIONS /Changing the Chart Display Method [Paging] sub-panel Used to paginate a chart for detailed viewing. Sets the number of horizontal Enables or disables the axis points displayed per page. change in scaling in the last page. Sets the current page number and total number of pages.
Page 464 APPLIED OPERATIONS /Changing the Chart Display Method [3D] sub-panel Change the zoom ratio. Used to set the 3D display. Rotates the chart in the horizontal direction with Switches between the 3D respect to the bottom side. and 2D chart display. Rotates the chart in the vertical direction with respect Sets the 3D chart display...
Page 465: Series] Panel
APPLIED OPERATIONS /Changing the Chart Display Method 2-15-2 [Series] Panel [Format] sub-panel Used to set the display method of a series of images. The set items are variable depending on the type of the chart. The following descriptions take a Line 3D chart and Point 3D chart as examples. This is the [Format] sub-panel when the Line 3D chart is selected.
Page 466 APPLIED OPERATIONS /Changing the Chart Display Method [Points] sub-panel Used to set the inflection points in the chart. Enables or disables the Sets the width of inflection inflection point display. points. Enables or disables the 3D Sets the height of inflection inflection point display.
Page 467 APPLIED OPERATIONS /Changing the Chart Display Method [Marks] sub-panel Used to set the items to be displayed along the coordinate points. Enables or disables the transparent background Enables or disables the item display. display in the chart. Enables or disables the item display of lines Sets the background color of outside the coordinate...
Page 468: Pop-Up Menus
APPLIED OPERATIONS /Pop-up Menus 2-16 Pop-up Menus Selection of function panels and display panels and other frequently-used FLUOVIEW functions (full-screen display, printer output, image save, LUT setting, comment setting) can be controlled by clicking the right button of the mouse, without selecting specific page tabs or buttons. Pop-up menu of function panel When the right button of mouse is clicked on the page tab of a function panel, a pop-up menu appears to allow selection of the function panel to be displayed at the front.
Page 469 APPLIED OPERATIONS /Pop-up Menus Pop-up menu of display panel When the right button of mouse is clicked on the page tab of a [Display] panel, a pop-up menu appears to allow selection of the [Display] panel to be displayed at the front. Click the mouse right button here.
Page 470 APPLIED OPERATIONS /Pop-up Menus Pop-up menu of comment When the right button of mouse is clicked on an comment in image, a pop-up menu appears to allow editing or deleting the specified comment. Fig. 2-156 Image Comment with Pop-up Menu Copies the comment.
Page 471 APPLIED OPERATIONS /Pop-up Menus Pop-up menu of image When the right button of mouse is clicked on the image, a pop-up menu appears to allow selection of image operations (full-screen display, printer output, image save, LUT setting, number of image divisions, comment editing).
Page 472 APPLIED OPERATIONS /Pop-up Menus Pop-up menu of the [Experiments in Memory] dialog box When the right button of mouse is clicked in the frame for opening an image file in the [File I/O], [Tile] or [Process] panel, a pop-up menu appears. This makes it possible to select the file to be opened when performing tile display, inter-image operations, image channel merger/extraction, etc.
Page 473: Appendix A List Of Tools
Appendix A List of Tools / List of Tools Appendix A List of Tools Appendix A-1 List of Tools Appendix A-1-1 Toolbar The horizontal array of buttons at the bottom left of the panel form the toolbar. The most frequently-used FLUOVIEW functions are gathered here. <Help>...
Page 474: Appendix A-1-2 Tools At The Top Of Display Panel
Appendix A List of Tools /List of Tools Appendix A-1-2 Tools at the Top of Display Panel These refer to the horizontal array of buttons at the top of the display panel. The functions frequently used with images are gathered here. <Set end position>...
Page 475: Appendix B List Of Hot Keys
Appendix B List of Hot Keys Appendix B List of Hot Keys Frequently-used FLUOVIEW functions (scanning, panel switching) can be controlled from the keyboard without using the mouse. Image acquisition-related keys Image acquisition channel selection Target Operation Cttrl + Switches the status of the Ch1 check box (Ch1 scanned/not scanned). Cttrl + Switches the status of the Ch2 check box (Ch2 scanned/not scanned).
Page 476 Appendix B List of Hot Keys Target Operation Completes image acquisition after completing series image acquisition. [Series Done] Stops scanning. [Stop Scan] Spave Scanning speed and area setting Target Operation Decreases the scan speed. Ctrl + Increases the scan speed. Ctrl + Decreases the zoom ratio.
Page 477 Appendix B List of Hot Keys Transmitted illumination lamp ON/OFF keys Target Operation Ctrl + Switches the transmitted illumination ON/OFF. <Trans. Lamp> Image Saving Target Operation Ctrl + Save the image in the [Display] panel being displayed as series images. <SAVE>...
Page 478 Appendix B List of Hot Keys [Acquire] sub-panels Target Operation Selects the [Settings] sub-panel. Alt + Selects the [Z Stage] sub-panel. Alt + Selects the [Time Series] sub-panel. Alt + Selects the [Dyes] sub-panel. Alt + Alt + Selects the [Lasers] sub-panel. Alt + Selects the [TIEMPO] sub panel .
Page 479: Appendix C Glossary
Appendix C Glossary Appendix C Glossary Clicking a command button with the mouse allows the function indicated on the button to be executed. Confocal AOTF Signifies the possibility of obtaining data on the plane AOTF represents Acoustic Optical Tunable Filter. where the irradiated laser beam is focused.
Page 480 Appendix C Glossary Double-click Action of pressing and releasing the mouse button quickly twice, without moving the moue. Icon An icon is a small figure with characters below it. The Drag icon indicates the status in which the window is Action of placing the mouse pointer on the target closed (or minimized).
Page 481 Appendix C Glossary Mouse A device which was named because it looks like a Resolution mouse. It is used to give instructions in the window. Number of dots composing the image on a screen or printer. When the number of dots is increased, i.e. Mouse pointer when the resolution is increased, the gradation can When the mouse is moved on the desk, arrow...
Page 482 Appendix C Glossary Text box A box in the window that accepts the input of character strings. Clicking a text box displays blinking |. This “|” indicates the position where the input character is inserted. Title bar The horizontal bar at the top of the window, that shows the title of the window or dialog box.
Page 483: Appendix D Formatting Of Magnetic Optical Disk
Appendix D Formatting of Magnetic Optical Disk Appendix D Formatting of Magnetic Optical Disk Use the Disk Administrator tool of Windows for formatting a magnetic optical (MO) disk. Note that this tool can be used only by a person who have the authority of Administrator. Click the <start>...
Page 484 Appendix D Formatting of Magnetic Optical Disk From the [Partition] menu of the [Disk Administrator] window, select the [Create] command. The dialog box as shown below appears. Set the value of the [Create partition of size] edit box in the [Create Primary Partition] dialog box to “Maximum size”, and click the <OK>...
Page 485 Appendix D Formatting of Magnetic Optical Disk 10. From the [Tool] menu of the [Disk Administrator] window, select the [Format] command. The dialog box as shown below appears. Fig. Appendix D-2 [Format] Dialog Box 11. Select [Removable Media (Unknown Size)] from the [Capacity] drop-down list in the [Format] dialog box. Then select [FAT] from the [File System] drop-down list and select [FAT] and select [Default Allocation Unit Size] from the [Allocation Unit Size] drop-down list.
Page 487: Using Excel
Appendix E Converting Analysis Data into a Chart Using EXCEL Appendix E Converting Analysis Data into a Chart Using EXCEL Start up Excel. From the [File] menu of Excel, select the [Open] command to open the analysis data file saved after analysis using FLUOVIEW. When the dialog box as shown below appears, click the [Delimited] option button in the [Original Data Type] group box, then select [Windows [ANSI]] from the [File Origin:] drop-down list.
Page 488 Appendix E Converting Analysis Data into a Chart Using EXCEL Click the <Next> button. When the dialog box as shown below appears, click the [General] option button in the [Columns Data Format] group box, then click the <Finish> button. Fig. Appendix E-3 Dialog Box Displayed When File is Opened with Excel (3/3) Drag the data to be converted into a chart.
Page 489 Appendix E Converting Analysis Data into a Chart Using EXCEL Click the <ChartWizard> button. <ChartWizard> button On the sheet, drag the area where the chart is to be inserted. The dialog box as shown below appears. Fig. Appendix E-4 Dialog Box of Chart Wizard (1/5) 9.
Page 490 Appendix E Converting Analysis Data into a Chart Using EXCEL 11. Select the desired chart format and click the <Next> button. The dialog box as shown below appears. Fig. Appendix E-7 Dialog Box of Chart Wizard (4/5) 12. Click the [Columns] option button under [Data Series in:]. 13.
Page 491 Appendix E Converting Analysis Data into a Chart Using EXCEL 17. Click the <Finish> button. A chart will be drawn on the sheet. Fig. Appendix E-9 Sheet with Inserted Chart When the mouse is clicked on the chart, black handles will appear around the chart. The following operations are available in this condition.
Page 492 Appendix E Converting Analysis Data into a Chart Using EXCEL When the mouse is double-clicked on the chart, a frame composed of oblique lines appear around the chart. (The frame may become a window when the chart is large.) The format settings are available in this condition. Click one of the following positions to display black handles.
Page 493: Appendix G-1 User Registration
Appendix F User Registration Appendix F User Registration Windows NT or Windows 2000 manages the system setting data on a per-user basis. Use the following procedure to register yourself as a user. Log in the system with Administrator. Register yourself as a new user. From the [Start] menu, select [Programs] - [Administrator] - [User Manager].
Page 494 Appendix F User Registration Enter the user name in the [Username] text box. It is not permitted to enter an existing user name or the same user name as the group name. The user name can be a character string of up to 20 characters and can contain any uppercase or lowercase characters except for the following characters.
Page 495: Appendix G User Registration Of Fv300
Appendix G USER REGISTRATION OF FV300 FLUOVIEW FV300 can store the system setup information (PMT Voltage, Gain, Offset, etc.) on a per-user basis. To make this possible, you have to register yourself as a user and log in personally when starting up the FV300 software.
Page 496 Appendix G USER REGISTRATION OF FV300/User Registration Click the <Add a user and Log in> button. The [New FLUOVIEW User] dialog box appears. [User Name:] text box Enter the user name to be registered. Enter the user name in the [User Name] text box and click the <OK> button. The newly set user name is added in the list box.
Page 497: Appendix G-2 Logging Into The Fv300
Appendix G USER REGISTRATION OF FV300/Logging into the FV300 Appendix G-2 Logging into the FV300 After the user name has been registered and the FV300 started, a dialog box for entering the user name appears. Enter the user name to log in the FV300. Double-click the [FLUOVIEW] icon on the desktop.
Page 498: Appendix G-3 Deleting A User
Appendix G USER REGISTRATION OF FV300/Deleting a User Appendix G-3 Deleting a User Double-click the [FLUOVIEW] icon on the desktop. The dialog box for entering the user name appears as shown below. [FLUOVIEW Setup] icon Enter “Administrator” in the [User Name:] text box and click the <OK> button. The [FLUOVIEW Setup] dialog box appears as shown below.
Page 499 Appendix G USER REGISTRATION OF FV300/Deleting a User Click the [Users] icon in [Hardware Settings] to display the [Users] panel at the front. [Users] icon <Add a user and Log in> button <Delete a user> button Used to add a new user. <Reset user to Factory Defaults>...
Page 500 Appendix G USER REGISTRATION OF FV300/Deleting a User Click the <Yes> button if you want to delete the user or the <No> button if you do not. The deleted user name disappears from the list. NOTE The user is deleted at the moment the <OK> button is clicked. Click the <Save New Setup>...
Page 501: Appendix H Change Of Default Folder For [File I/O] Panel
Appendix H Change of Default Folder for [File I/O] Panel FLUOVIEW FV300 usually opens the default folder for the [File I/O] panel (C: FLUOVIEW IMAGES) to save acquired images or load saved images. The default folder for saving an image can be changed or a desired folder can be specified directly when loading a saved image.
Page 502 Appendix H Change of Default Folder for [File I/O] Panel Click the [Start] button to open the [Start] menu. Then select commands [Settings] - [Control Panel] - [System]. The [System Properties] dialog box appears as shown below. OPERATION INSTRUCTIONS IV . Page H - 2...
Page 503 Appendix H Change of Default Folder for [File I/O] Panel Select the [Environment] sub-panel. User name logging in Windows NT or Windows 2000. Select [FLUOVIEWIMAGES] under [Variable] in the [User Variables for Administrator:] list box. “Administrator” of [User Variables for Administrator:] is variable depending on the user name logging in Windows NT or Windows 2000.
Page 504 Appendix H Change of Default Folder for [File I/O] Panel In the [Value:] text box, enter the path name of the default folder to be changed by delimiting the drive and folder names using backslashes ( ). (The figure shows an example in which folder IMAGE in drive C is entered.) [Value:] text box Enter the path name of the default folder.
Page 505 Appendix H Change of Default Folder for [File I/O] Panel Click the <OK> button to close the [System Properties] dialog box. Now, the default folder for the [File I/O] panel will be changed from the next time you start FLUOVIEW. When the default folder is changed to C: IMAGES New default folder C IMAGES...
Page 507: Appendix I List Of Functions In The [Active Overlays] Dialog Box
Appendix I List of Functions in the [Active Overlays] Dialog Box /Coordinate Position Data Appendix I List of Functions in the [Active Overlays] Dialog Box Active Overlays is a kind of overlay function displayed on an image. Active Overlays does not simply show the entered characters, but searches the image data related to the keyword specified in <...
Page 508: Appendix I-1-2 Y-Coordinate
Appendix I List of Functions in the [Active Overlays] Dialog Box /Coordinate Position Data Appendix I-1-2 Y-Coordinate The Y-coordinate position with respect to the top left corner of the screen is displayed. «Syntax» <y[ [hotspot] [raw/calibrated] value [units]]> Arguments inside [ ] can be omitted. If [raw/calibrated] is omitted, the same setting as when [calibrated] is specified will be applied.
Page 509: Appendix I-1-3 Other
Appendix I List of Functions in the [Active Overlays] Dialog Box /Coordinate Position Data Appendix I-1-3 Other Positions can also be displayed by entering the following keywords in the syntax in place of the X- and Y-coordinate positions described in sections J-1-1 and J-1-2. 1 Z Position With cross-section related images such as XYZ images, the Z position with respect to the first image can be displayed.
Page 510: Appendix I-2 Intensity Data
Appendix I List of Functions in the [Active Overlays] Dialog Box /Intensity Data Appendix I-2 Intensity Data The intensity value can be displayed. When images are overlapped, the intensity value of each image is accompanied with the channel number placed after it. «Syntax»...
Page 511: Appendix I-3 Other
Appendix I List of Functions in the [Active Overlays] Dialog Box Appendix I-3 Other The following image data can be displayed in addition to the X- and Y-coordinate positions and intensity value. Appendix I-3-1 Channel Number The channel number can be displayed. When images are overlapped, the displayed channel numbers are connected by the “+”...
Page 513: Appendix J Hand Switch And Microscope Frame Function Allocation
J Hand Switch and Microscope Frame Function Allocation/ Appendix Hand Switch Functions Appendix J Hand Switch and Microscope Frame Function Allocation When the U-HSTR2 hand switch, the BX51/BX61/IX81 microscope frames, and U-FH focus adjustment knob are shipped from the factory, the following functions have been allocated to their control buttons. These function allocations are valid only when the FLUOVIEW NOTE software is started.
Page 514: Appendix J-2 Microscope Frame Functions
J Hand Switch and Microscope Frame Function Allocation/ Appendix Microscope Frame Functions Appendix J-2 Microscope Frame Functions Appendix J-2-1 BX Left side Fig. Appendix J-2 BX Microscope Frame Button Function Stage escape/return switching Focus fine/coarse adjustment switching Z position upper limit setting in Z-observation Z position lower limit setting in Z-observation Excitation DM position up Excitation DM position down...
Page 515: Appendix J-2-2 Ix
J Hand Switch and Microscope Frame Function Allocation/ Appendix Microscope Frame Functions Appendix J-2-2 IX Front Left side Fig. Appendix J-3 IX81 Microscope Frame Button Function Focus fine/coarse adjustment switching Excitation DM position up Excitation DM position down BI/LSM light path switching Objective revolving nosepiece position up Objective revolving nosepiece position down Transmitted light (LG-PS2) ON/OFF switching...
Page 516: Appendix J-2-3 Focus Adjustment Knob
J Hand Switch and Microscope Frame Function Allocation/ Appendix Microscope Frame Functions Appendix J-2-3 Focus Adjustment Knob 1 BX Fig. Appendix J-4 Focus Adjustment knob (BX) Button Function Z position upper limit setting in Z-observation Z position lower limit setting in Z-observation Excitation DM position up Excitation DM position down Focus fine/coarse adjustment switching...
Page 517 On This Volume This volume describes the user maintenance procedures of the FLUOVIEW FV300 system. Please read this volume so that you can use the system for an extended period of time.
Page 519 CONTENTS 1 Software Setup 1-1 Re-setup or Updating of the Software ........... 1-1 1-2 New Setup of the Software ............. 1-3 1-3 Setting the System Configuration..........1-6 1-3-1 Overall Setting of FLUOVIEW ..............1-6 1-3-2 Setting the [Microscope Control Panel] ............1-14 1-1 Adding the dyeing method ............
Page 521: Software Setup
Software Setup/ Re-setup or Updating of the Software 1 Software Setup The FLUOVIEW software has been set up before it is delivered to the user. A CD-ROM containing the software program is provided with FLUOVIEW. This CD-ROM is intended for use when re-setup is required due to a fault or when the user wants to set up the software newly.
Page 522 Software Setup/ Re-setup or Updating of the Software Continuously, deletion of software is started. For deletion, carry out according to a wizard. Continuously, installation of the software of a new version is started. Refer to 1-2 “New Setup of the Software” for details. MAINTENANCE Page 1 - 2...
Page 523: New Setup Of The Software
Software Setup/ New Setup of the Software 1-2 New Setup of the Software The following procedure allows you to set up the FLUOVIEW software in a computer you use. Load the FLUOVIEW setup CD-ROM in the CD-ROM drive. The dialog box as shown below will appear. If the dialog box doesn’t appear, run the 'Install.exe' file that is present in the root directory of the CD-ROM.
Page 524 Software Setup/ New Setup of the Software When the [Choose Destination Location] dialog box appears, confirm the setup destination drive name and directory and select the <Next> button. MAINTENANCE Page 1 - 4...
Page 525 Software Setup/ New Setup of the Software When the setup has completed, the [Setup Complete] dialog box appears. Select the [Yes, I want to restart my computer now.] option button and press the <Finish> button. This will restart the computer. MAINTENANCE Page 1 - 5...
Page 526: Setting The System Configuration
[FLUOVIEW Setup] icon Enter the user name in the [User Name:] text box and click the <OK> button to log into FLUOVIEW FV300. For details, see Appendix G-2, “Logging into the FV300”. The dialog as shown below appears for use in saving the system configuration.
Page 527 Software Setup/ Setting the System Configuration Click the [Z Stage] icon in [User Settings] to display the [Z Stage] panel in the front position, and check and set the presence of motor controller, positioning of the motor controller, backlash and moving amount of the Z motor. Set the backlash of motor operation in the [Z Stage] sub-panel.
Page 528 Software Setup/ Setting the System Configuration Click the [Objectives] icon in [User Settings] to display the [Objectives] panel in the front position. In this panel, select the items to be included in the list of objectives used by the FLUOVIEW application. (Each user should set the objectives that the user wants to use with FLUOVIEW.) [Objectives] icon <To delete an unnecessary objective from the list>...
Page 529 Software Setup/ Setting the System Configuration Click the [Fluorescence] icon in [User Settings] to display the [Fluorescent Deys/Colors panel in the front position. In this panel, select the items to be included in the list of dyes used by the FLUOVIEW application.
Page 530 Software Setup/ Setting the System Configuration <To change the button settings> Press the button to indicate that you want to change the setting. When the dialog box as shown below appears, specify the button name and LUT file name and select the <OK>...
Page 531 Software Setup/ Setting the System Configuration Display the [Hardware] panel in the front position. In this panel, select Enabled/Disabled to enable/disable the automatic black-level adjustment, enter the values for delay timing for TD unit, and enter the maximum duration for continuous bi-directional scanning when setting fast scan mode. [Delay Timing for TD Unit] group box This data should usually be left in the default values and does not have to be changed.
Page 532 Software Setup/ Setting the System Configuration 10. Click the [Lasers] icon in [Hardware Settings] to display the [Lasers] panel in the front of position. Set the lasers to be used and set its default intensity. [Default Laser Intensity] scale Set the initial ND filter value at the start [Lasers Installed] check box of the FLUOVIEW application.
Page 533 Software Setup/ Setting the System Configuration 12. The [Registration] panel have been set at the factory and do not need to be changed here. 13. After completing the setup, select the <Save New Setup> button on the bottom left of the panel. (Selecting <Quit w/o Saving>...
Page 534: Setting The [Microscope Control Panel]
Software Setup/ Setting the System Configuration 1-3-2 Setting the [Microscope Control Panel] The <Microscope Setting> button as shown below appears when selecting “BX 51,52”, “BX51,52WI”, or “IX71” and checking the [Automatic] check box or selecting “BX61,62”, “BX61,62WI”, or “IX81” in the [Microscope] group box in the [Microscope] panel in the [FLUOVIEW Setup] dialog box.
Page 535 Software Setup/ Setting the System Configuration [Shutter] group box Sets up to use the shutter for transmitted observation. [Filter Turret] group box Sets up the name and color of the filter turret for transmitted o Select the initial status of display magnification when multiple images are to be displayed by displaying multiple [Display] panels together.
Page 536 Software Setup/ Setting the System Configuration The example of setting procedure in the [Microscope Setup] window. Changing the Number of Buttons Section 1-3-1-1 Setting the Button Section 1-3-1-2 Editing the name of the buttons in the [Mirror Unit], [Condenser Turret] and [Filter Turret] group boxes Setting the Name Editing in the [Nosepiece] group box Setting the Name...
Page 537 Software Setup/ Setting the System Configuration 1-3-2-1 Setting the Number of Buttons Right-click the mouse on the area outside of the buttons in the group box where the number of buttons to be displayed is changed. The pop-up menu as shown below appears.
Page 538 Software Setup/ Setting the System Configuration Select “Edit”. The [Edit] dialog box as shown below appears. [Magnification] text box [Recommended Confocal Pinhole] text box Enter the magnification of Enter the number of the confocal pinhole with keyboard. the objective with keyboard.
Page 539 Software Setup/ Setting the System Configuration 2 Setting the color of the button engaged into the light path In [Microscope Control Panel] window on the software, specify the color of the button of the cube, the objective, and the condenser engaged into the light path. Right-click the mouse on the area outside of the buttons in the [Mirror Unit], [Nosepiece], [Condenser Turret], or [Filter Turret] group box.
Page 540 Software Setup/ Setting the System Configuration 1-3-2-4 Finishing the setting Finishing the setup and closing the [Microscope Setup] window. <Quit w/o Saving> button <Save New Setup> button Click the < Save New Setup> button to save the setup and close the window. Or click the <Quit w/o Saving>...
Page 541: Adding The Dyeing Method
Software Setup/ Adding the dyeing method 1-4 Adding the dyeing method Newly add a dyeing method and set laser type, excitation wavelength, and emission wavelength. This section describes a simple example of adding CFP as a dyeing method and setting laser type, excitation wavelength, and emission wavelength.
Page 542 Software Setup/ Adding the dyeing method Display the [Fluorescent Dyes/Colors] panel at the front position in the [FLUOVIEW Setup] dialog box. Double-click here to add the dyeing method. Double-click the “Double click here to make a new Dye” in the [Dyes on your Samples] list.
Page 543 Software Setup/ Adding the dyeing method Enter the value of the emission wavelength into the [WaveLength 1] text box in the [Emission Wave Length] group box (e.g. 480). photometry, enter the value of the wavelength into the [WaveLength 2] text box too.
Page 544: Associating A Detection Channel And Filter To The Dyeing Method
Software Setup/ Adding the dyeing method 1-4-1 Associating a Detection Channel and Filter to the Dyeing Method The detection channel and filter can be associated to a dyeing method using the [Optical System Configuration Setup] dialog box. Display [FLUOVIEW Setup] dialog. See section 1-3-1 of this manual to display [FLUOVIEW Setup] dialog.
Page 545 Software Setup/ Adding the dyeing method Click [Edit Dye Assign Database] button. [Optical System Configuration Setup] dialog box appears as shown below. [Barrier Filter] drop-down list [Beam Splitter] drop-down list Select the barrier filter for Select the beam splitter for each channel. each channel.
Page 546 Software Setup/ Adding the dyeing method Associations between dyeing methods and detection channels are saved per dyeing method combination, and applied when the combination is selected in the [Dyes] sub-panel of the FLUOVIEW software. Associations between dyeing methods and filters are saved per dyeing method combination, and displayed as setting guide on [Microscope configuration] window when the said combination is selected in the [Dyes] sub-panel of the FLUOVIEW software.
Page 547 Software Setup/ Adding the dyeing method the blank. In this example, select CFP for channel 1. After completing the above, click the <Save> button to close the [Optical System Configuration Setup] dialog box. MAINTENANCE Page 1 - 2 7...
Page 549: Maintenance Of Major System Units
Maintenance of Major System Units /Scan Unit 2 Maintenanc e of Major System Units This section describes the maintenance of the following units. Scan unit Laser scanning microscope 2-1 Scan Unit The scan unit used with this system has been designed to allow replacement of the BARRIER FILTER slider and DETECTION MODE slider in order to accommodate a wide range of research.
Page 550: Laser Scanning Microscope
Maintenance of Major System Units/ Laser Scanning Microscope 2-2 Laser Scanning Mi croscope According to IEC60825 “Safety of Laser Products” and EN60825, this product is classified as a CLASS 3B laser product. However, as it belongs to “CLASS 3B with a laser wavelength range from 400 to 700 nm, which is no more than 5 times the AEL of CLASS 2”, all of the microscope modules listed below can be attached or removed as a part of their maintenance activities.
Page 551 Maintenance of Major System Units/ Laser Scanning Microscope The definitions of the term “maintenance” by EN60825 and CDRH 21CFR are quoted below for reference. IEC60825(EN60825) Maintenance: The performance of those adjustments or procedures specified in user information provided by the manufacturer with the laser product, which are to be performed by the user for the purpose of assuring the intended performance of the product.
Page 553 On This Volume This volume describes the treatment against possible troubles. In case of trouble, please read volume before calling for service. If the normal operation cannot still be restored, please contact your local Olympus representative.
Page 555 CONTENTS 1 TROUBLESHOOTING GUIDE 1. Fluorescence image cannot be observed........1-1 2. Transmitted image cannot be observed.......... 1-2 3. Image is disturbed................1-2 4. Image is striped.................. 1-2 5. Image looks poor................1-2 6. Image is irregularly blurred or the brightness is uneven....1-3 7.
Page 557: Troubleshooting Guide
If you cannot solve the problem after checking the entire list, please contact your local Olympus representative for assistance. Before contacting Olympus, please fill the “Inquiry Table” at the end of this volume and inform Olympus of its contents. Phenomenon...
Page 558: Fluorescence Image Cannot Be Observed
OPERATION the microscope. INSTRUCTIONS 3. Image is disturbed. The installation location is Consult Olympus. subject to large vibrations. External light from a Darken the room before fluorescent lamp, etc. is acquiring image. detected.
Page 559: Image Is Irregularly Blurred Or The Brightness Is Uneven
When dirt cannot be taken, moisten the gauze with a slight amount of 3:7 alcohol:ether mixture or benzine. The installation location is Consult Olympus. subject to large vibrations. 6. Image is irregularly blurred The specimen is tilted. Set the specimen Instruction manual or the brightness is horizontally.
Page 560: The Position Reproduction Of The Z-Motor Is Poor
TROUBLESHOOTING GUIDE Phenomenon Cause Treatment Manual Ref. Pages 10. Image is dark and noisy. The confocal aperture is too Adjust the confocal aperture 1-2-6, OPERATION small. to an optimum size. INSTRUCTIONS The HV of photomultiplier Set HV at no more than 800. 2-2-1-3-9, tube exceeds 800.
Page 561: Fluoview Software Cannot Be Started
Then exit from windows and reboot the computer. 19. The power supply of The fuse has run out. Contact your local Olympus Power Unit(FV5-PSU) representative for assistance is not turned on. then replace the fuse. 20. Bios some check error...
Page 564 2 Corporate Center Drive, Melville, NY 11747-3157, U.S.A. OLYMPUS SINGAPORE PTE LTD. 491B River Valley Road, #12-01/04 Valley Point Office Tower, Singapore 248373 OLYMPUS OPTICAL CO. (U.K.) LTD. 2-8 Honduras Street, London EC1Y OTX, United Kingdom. OLYMPUS AUSTRALIA PTY. LTD.

Olympus FLUOVIEW FV300 User Manual

1 System Overview 1-1

1 Getting Started Fluoview 1-1

Getting Started FLUOVIEW

Applied Operations

Appendix A List of Tools

Appendix A-1 List of Tools

Appendix A-1-1 Toolbar

Appendix A-1-2 Tools at the Top of Display Panel

Appendix B List of Hot Keys

Appendix C Glossary

Appendix D Formatting of Magnetic Optical Disk

Appendix E Converting Analysis Data into a Chart

Appendix G-1 User Registration

Appendix F User Registration

Appendix G USER REGISTRATION of FV300

Appendix G-2 Logging into the FV300

Appendix G-3 Deleting a User

Appendix H Change of Default Folder for [File I/O] Panel

Appendix I List of Functions in the [Active Overlays] Dialog Box

Appendix I-1 Coordinate Position Data

Appendix I-1-1 X-Coordinate

Appendix I-1-2 Y-Coordinate

Appendix I-1-3 Other

Appendix I-2 Intensity Data

Appendix I-3 Other

Appendix I-3-1 Channel Number

Appendix I-3-2 Objective Power

Appendix I-3-3 Date of Image Capturing

Appendix I-3-4 Time of Image Capturing

Appendix I-3-5 Image File Name

Appendix J Hand Switch and Microscope Frame Function Allocation

Appendix J-1 Hand Switch Functions

Appendix J-2 Microscope Frame Functions

Appendix J-2-1 BX

Appendix J-2-2 IX

Appendix J-2-3 Focus Adjustment Knob

1 Troubleshooting Guide 1-1

8 Circular Flare Is Observed at the Center of Image

9 Image Is Blurred or out of Focus

Quick Links

Chapters

Related Manuals for Olympus FLUOVIEW FV300

Summary of Contents for Olympus FLUOVIEW FV300