Page 1: Quick Start
FV1000 LASER SCANNING BIOLOGICAL MICROSCOPE FV10-ASW [Ver2.0] Notice Thank you for your purchase of Olympus microscope at this time. AX7797 Hold this manual by your side when using this microscope all the time and keep it with care after reading.
Page 3 Caution Caution FV1000MPE is a CLASS 4 laser product; FV1000 is a CLASS 3B laser product. The procedures for using this system are classified as follows: Service “Service” means any adjustment or repair performed by highly trained and skilled technical personnels who are provided the service training following to the service manual for this system.
Page 4 In case of purchasing the laser simultaneously, we have prepared the following manual for the laser. MaiTai Series User’s Manual [Quick Start] In addition, we have prepared one service manual for the laser as below. Technical personnels who perform the laser service require to take the service training.
Page 5 CONTENTS This Quick Start has divided into the volume by the following system configurations. IX81 (Spectral Type) IX81 (Filter Type) BX61 (Spectral Type) BX61 (Filter Type)
Page 7 Laser Conforcal Scanning Microscope FV1000D Spectral Type (invertedMicroscpeIX81) Operation Manual...
Page 8: Table Of Contents
Contents Contents System introduction FV1000D Laser DyeList System Preparation Visible Observation Observation of Fluorescence image Observation of Differential Interference Contrast Images Image Acquisition Overview of Operation Panel for Image Acquisition Single Stain on XY Image 9-11 Complement of adjusting the image 12-13 Double Stain on XY Image 14-15...
Page 9 Spectral Type Main Scanner...
Page 10 Dye List (FV1000D Lasers are available below) LD405nm LD440nm LD473nm LD559nm LD635nm Ar458nm Ar488nm Ar515nm HeNe(G)543nm...
Page 11: System Preparation
System Preparation System Preparation 1. Turn the computer ON. [In case of equipped concentrated power supply, power on it first] 2. Turn the laser ON (Turning the key switch) 2-1. LD559nm ON 2-2. Multi Ar 458nm 488nm 515nm 2-2. HeNe(G)(643nm) ON 3.
Page 12: Observation Of Fluorescence Image
Visual Observation under the Microscope Visual Observation under the Microscope Observation of Fluorescence Image 1. Select an objective lens by using the hand switch Hand switch 2. Select florescent filter cube MEMO Fluorescence filter NIBA: Blue Excitation / Green Fluorescence (Ex.:FITC,EGFP) WIG: Green Excitation / Red Fluorescence (Ex.:Rhodamine, DsRed)
Page 13: Observation Of Differential Interference Contrast Images
Visual Observation under the Microscope Visual Observation under the Microscope Observation of Differential Interference Contrast Images 1. Select the Objective Lens Hand switch *DIC prism 2. Insert the Polarizing Plate in the Light Pass 3. Insert the DIC prism slider in the light pass Click the button on Fluoview software...
Page 14: Overview Of Operation Panel For Image Acquisition
Overview of Operation Panel for Image Acquisition Overview of Operation Panel for Image Acquisition Scan mode Scan speed Number of pixels Zoom & Pan Laser output adjustment Objective lens Focus Time Interval & Time Number (for acquisition of XYT or XT image) Transmitted light observation (visual observation)
Page 15: Single Stain On Xy Image
Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Single stain of green fluorescence dye (FITC) 1. Click on the FV10-ASW software button to close the fluorescence lamp shutter.
Page 16 Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) 4. Press XY Repeat button click to get image Continuous scan mode 5. Focus to the specimen 6. Adjust the green (FITC) image. Adjust sensitivity of HV and ·...
Page 17 Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) Select AutoHV and then select ScanSpeed. *As the scan speed becomes slower, noise can be removed while maintaining the current brightness. Press the Stop button to stop scanning Click on XY, and "2D View-LiveImage(x)"...
Page 18: Complement Of Adjusting The Image
Complement of adjusting the image Complement of adjusting the image 1. Click “Clip scan” button , and enclose an interesting region’ s image on the whole image. 2. pixel setting * The standard pixel is 512 x 512 3. Zoom Setting Press “XY Repeat”...
Page 19 Complement of adjusting the image Complement of adjusting the image 5. Pan X,Y Be able to move the field of view to set Pan X,Y without stage action 6. Rotation PanX,Y and Rotation reset button Be able to rotate the whole image. Click “Auto”...
Page 20: Double Stain On Xy Image
Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Double stain of green fluorescence dye (Alexa 488) and red fluorescence dye (Alexa 546) Simultaneous scan 1.
Page 21 Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) 4. Press the XY Repeat button to start scanning. 5. Adjust the green (Alexa Fluor 488) image and the red (Alexa Fluor 546) image. (The image adjustment is outlined below. For more information, refer to Appendix 1.) 6.
Page 22: Sequential Scan Line Sequential
Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Double stain of green fluorescence dye (Alexa 488) and red fluorescence dye (Alexa 546) Sequential scan (Line Sequential is introduced here.) 1.
Page 23: Double Stain On Xyz Image
Image Acquisition Image Acquisition (Double Stain on XYZ Image) (Double Stain on XYZ Image) Acquisition of 3D images (XYZ) (fluorescence image only) Sample: Double stain of green fluorescence dye (FITC) and red fluorescence dye (Rhodamine) This is the procedure to acquire images through Line Sequential scanning.
Page 24 Image Acquisition Image Acquisition (Double Stain on XYZ Image) (Double Stain on XYZ Image) 9. Select AutoHV and then select ScanSpeed. 10. Select Depth. 11. Press the XYZ button to acquire an image. 12. Click on “SeriesDone”, and "2D View-LiveImage(x)" is displayed on the window bar for the image that has been acquired.
Page 25: Four Stain On Xy Image
Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) Acquisition of 4 stain images (XY) (fluorescence image only) Sample: Four stain of Blue fluorescence dye (DAPI) ,green fluorescence dye (Alexa488) and red fluorescence dye (Rhodamine), far-red fluorescence dye (Cy5) This is the procedure to acquire images through Virtual Channel scan Select Virtual channel...
Page 26 Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) 4. Select “Phase1”, “DAPI”, Alexa488, RhodaminRed are registered on ImageAquisitionControl. * Slit and Filter, DM are automatically set for ”DAPI”, “Alexa488” or ”PhodaminRed” 5. Select “Phase2”, Cy5 is registered on ImageAquisitionControl * Slit and Filter, DM are automatically set for ”Cy5”...
Page 27 Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) 7. Click on Virtual Channel Controller to acquire the image. * Be able to start at each Phase. 8. Saving the image: Right-click on the Image Icon displayed on the Data 8.
Page 28: Single Stain + Dic On Xy Image
Image Acquisition Image Acquisition (Single Stain + DIC on XY Image) (Single Stain + DIC on XY Image) Acquisition of a single image (XY plane) (fluorescence image and differential interference contrast image) Sample: Green fluorescence dye (FITC) and differential interference contrast image 1.
Page 29 Image Acquisition Image Acquisition (Single Stain + DIC on XY Image) (Single Stain + DIC on XY Image) 5. Press the “XY Repeat” button to start scanning. 6. Adjust the green (FITC) image and the differential interference contrast image. 7. Press the “Stop button” to stop scanning.
Page 30: Merge The Image Between Fluorescent Xy Image And Dic Image
Merge the images between Merge the images between fluorescent XY image and image fluorescent XY image and image Edit different each files to the same file. This is available for making merge image Between fluorescent image and focused DIC image. 1.
Page 31 Image Acquisition Image Acquisition (Single Stain on XYZT Image) (Single Stain on XYZT Image) This is available for the Time series scan experiment. 1. Adjust the image. * Refer P17,18 2. Enter interval time to “Interval” Enter interval number to “Num” Example: Acquiring time series scan images every 5minutes for 1hour is below, 3.
Page 32 Image Acquisition Image Acquisition (Single Stain on XYZT Image) (Single Stain on XYZT Image) 1. Adjust the image. * Refer P17,18 2. Insert ZDC unit to left side. 3. Check “EnableZDC AF during Time Series Scan” and click “ZDC setting”. 4.
Page 33 Image Acquisition (Spectral Image on XYL Image) Image Acquisition (Spectral Image on XYL Image) Acquisition of a spectral image (XYL) Sample: Double stain of green fluorescence dye (Alexa Fluor 488) and green fluorescence dye (YOYO-1) 1. Click on the FV10-ASW software button to close the fluorescence lamp shutter.
Page 34 Image Acquisition (Spectral Image on XYL Image) Image Acquisition (Spectral Image on XYL Image) Click on the button, and the Spectral Setting window appears. Set the slit width for CHS1 to 20 nm, for example. Press the XY Repeat button to start scanning.
Page 35 Image Acquisition (Spectral Image on XYL Image) Image Acquisition (Spectral Image on XYL Image) 10. Set the range of wavelength to be acquired, the slit width and the step. Start = Start wavelength End = End wavelength Resolution = Slit width StepSize = Step Select AutoHV and then select ScanSpeed.
Page 36 Image Analysis ( Unmixing Image Analysis ( Unmixing I. When each fluorescence dye point is clear I. When each fluorescence dye point is clear From an XYL image where fluorescence dyes with similar fluorescence spectrums are present together, derive the fluorescence spectrum for each fluorescence dye and obtain an unmixed image based on the fluorescence spectrums.
Page 37 Image Analysis ( Unmixing Image Analysis ( Unmixing I. When each fluorescence dye point is clear I. When each fluorescence dye point is clear Sample: single stain of green fluorescence dye (GFP) and auto fluorescence from cell 1. Open the XYL image (GFP + auto fluorescence).
Page 38: Unmixing
Image Analysis ( Unmixing Image Analysis ( Unmixing II. When a control sample is used II. When a control sample is used From an XYL image with a single type of fluorescence dye, derive the fluorescence spectrum of the dye and obtain an unmixed image based on the fluorescence spectrum.
Page 39: Image Analysis
Image Analysis ( Unmixing Image Analysis ( Unmixing III. When only the number of types of fluorescence dyes is known III. When only the number of types of fluorescence dyes is known (Blind Unmixing (Blind Unmixing From an XYL image where fluorescence dyes with similar fluorescence spectrums are present together, obtain an unmixed image based on only the number of types of fluorescence dyes.
Page 40 Reload the image conditions Reload the image conditions 1. Open the file and click 2. Click 3. The conditions (HV,Offset, CA and so on ) are reloaded .
Page 41 Overview of the 2D Operation Panel Overview of the 2D Operation Panel Display switching Click on the button to switch the display Frame advance Enlarge 1:1 display Animation Adjust to the Ch1 display window size Advance speed TEXT Ch2 display Various kinds of ROIs formulation...
Page 42 Image Analysis (Acquire a Projection Images) Image Analysis (Acquire a Projection Images) 1. Click on the button to select 2. To save this image, right-click on the image, select Save Display and save the image with a new name.
Page 43 Image Analysis Image Analysis (Save a Z section Image as 2D file) (Save a Z section Image as 2D file) Save the image in step 3 or 5 6. Click on the button. 7. A 2D View-(file name) image is created.
Page 44 Image Analysis (Inserting the Scale Bar) Image Analysis (Inserting the Scale Bar) 1. Click on the button. 2. While left-clicking the image, drag and drop it at a certain point. Change the size 3. While clicking the right or left handle, move the mouse from side to side.
Page 45 Image Analysis Image Analysis (Rotating a Three dimensional Image) (Rotating a Three dimensional Image) 1. Click on the button for a 2D View-(file name) image. 2. A 3D view is created. 3. Drag the mouse on the image to observe it at a certain angle. Simple animation 4.
Page 46 Image Analysis (Saving an Image) Image Analysis (Saving an Image) Convert each channel of an XY or XYZ image into a TIFF format 1. Right-click on the Image Icon displayed on the Data Manager and select Export. 2. Set Save as type to TIFF. 3.
Page 47 Image Analysis Image Analysis (Rotating a Three dimensional animation) (Rotating a Three dimensional animation) To save a rotation file as an animated image, create three- dimensional images according to the following procedure. For example, try to rotate an image by 180 degrees.
Page 48: Edit The Image Color And Contrast
2D Image Analysis 2D Image Analysis ( Edit the image color and contrast) ( Edit the image color and contrast) 1. Click “LUT” and then LUT table appears below, 2.Edit contrast to drag to left or right side, and another way to 3.
Page 49: The Image Of Z Section
2D Image Analysis 2D Image Analysis ( the image of Z section ) ( the image of Z section ) 1. Click and select again, then Projection image is shown on 2D View after getting XYZ image. 2. Click and select 3.
Page 50: Intensity Profile Of Each Z Sections
2D Image Analysis 2D Image Analysis ( Intensity Profile of each Z sections ) ( Intensity Profile of each Z sections ) 1. Click and then 2D Control Panel is shown below, 2. Click “Profile” and then Intensity Profile of each Z sections is shown on the X and Y axis.
Page 51: Measure
2D Image Analysis 2D Image Analysis ( Measure ) ( Measure ) 1. Enclose interesting regions by ROI Line on interesting positions by ROI 2. Click “measure”. 4. According to click “Measure All ROIs” , then the information of all ROI is calculated on Region Measurement.
Page 52 2D Image Analysis 2D Image Analysis ( Line Intensity Profile on the 2D image ) ( Line Intensity Profile on the 2D image ) 1. Line on the 2D image by ROI 2. Click “Intensity Profile” 3. “Intensity Profile” on the line is shown as intensity graph .
Page 53: Line Series Analysis / Co-Localization
2D Image Analysis 2D Image Analysis Line Series Analysis Line Series Analysis 1. Line on the 2D image. 2. Click “Line Series Analysis” 3. Intensity of Z position/ time on the line is shown as a graph . 2D Image Analysis 2D Image Analysis ( Co localization )
Page 54 2D image Analysis (Series Analysis TimeLapse) 1. Enclose interesting regions by ROI 2. Click “Series Analysis” 3. “Series Analysis” graph is shown below, Y axis shows intensity, X axis shows time and then be able to see time series reaction each ROIs.
Page 55: Closing The System
Closing the System Closing the System 1. Exit the FV10-ASW software by selecting File/Exit. 2. Exit the Windows. (1) Select Start/Shut Down. (2) On the Shut Down Window, select Shut Down and click on OK. 3. Turn the laser OFF. (Turn the key switch to the OFF position.) 3-1.
Page 57 Laser Conforcal Scanning Microscope FV1000D Filter Type (invertedMicroscpeIX81) Operation Manual...
Page 58 Contents Contents System introduction FV1000D Laser DyeList System Preparation Visible Observation Observation of Fluorescence image Observation of Differential Interference Contrast Images Image Acquisition Overview of Operation Panel for Image Acquisition Single Stain on XY Image 9-11 Complement of adjusting the image 12-13 Double Stain on XY Image 14-15...
Page 59 Filter Type Main Scanner...
Page 60 Dye List (FV1000D Lasers are available below) LD405nm LD440nm LD473nm LD559nm LD635nm Ar458nm Ar488nm Ar515nm HeNe (G) 543nm...
Page 61: System Preparation
System Preparation System Preparation 1. Turn the computer ON. [In case of equipped concentrated power supply, power on it first 2. Turn the laser ON (Turning the key switch) 2-1. LD559nm ON 2-2. Multi Ar 458nm 488nm 515nm 2-2. HeNe(G) (643nm) ON 3.
Page 62: Observation Of Fluorescence Image
Visual Observation under the Microscope Visual Observation under the Microscope Observation of Fluorescence Image 1. Select an objective lens by using the hand switch Hand switch 2. Select florescent filter cube MEMO Fluorescence filter NIBA: Blue Excitation / Green Fluorescence (Ex.:FITC,EGFP) WIG: Green Excitation / Red Fluorescence (Ex.:Rhodamine, DsRed)
Page 63: Observation Of Differential Interference Contrast Images
Visual Observation under the Microscope Visual Observation under the Microscope Observation of Differential Interference Contrast Images 1. Select the Objective Lens Hand switch *DIC prism 2. Insert the Polarizing Plate in the Light Pass 3. Insert the DIC prism slider in the light pass Click the button on Fluoview software...
Page 64: Overview Of Operation Panel For Image Acquisition
Overview of Operation Panel for Image Acquisition Overview of Operation Panel for Image Acquisition Scan mode Scan speed Number of pixels Zoom & Pan Laser output adjustment Objective lens Focus Time Interval & Time Number (for acquisition of XYT or XT image) Transmitted light observation (visual observation)
Page 65: Single Stain On Xy Image
Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Single stain of green fluorescence dye (FITC) 1. Click on the FV10-ASW software button to close the fluorescence lamp shutter.
Page 66 Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) 4.Press XY Repeat button click to image Continuous scan mode 5. Focus to the specimen 6. Adjust the green (FITC) image. Adjust sensitivity of HV and ·...
Page 67 Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) Select AutoHV and then select ScanSpeed. *As the scan speed becomes slower, noise can be removed while maintaining the current brightness. Press the Stop button to stop scanning Click on XY, and "2D View-LiveImage(x)"...
Page 68: Complement Of Adjusting The Image
Complement of adjusting the image Complement of adjusting the image 1. Click “Clip scan” button , and enclose an interesting region’s image on the whole image. 2. pixel setting * The standard pixel is 512 x 512 3. Zoom Setting Press “XY Repeat”...
Page 69 Complement of adjusting the image Complement of adjusting the image 5. Pan X,Y Be able to move the field of view to set Pan X,Y without stage action 6. Rotation PanX,Y and Rotation reset button Be able to rotate the whole image. Click “Auto”...
Page 70: Double Stain On Xy Image
Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Double stain of green fluorescence dye (Alexa 488) and red fluorescence dye (Alexa 546) Simultaneous scan 1.
Page 71 Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) 4. Press the XY Repeat button to start scanning. 5. Adjust the green (Alexa Fluor 488) image and the red (Alexa Fluor 546) image. (The image adjustment is outlined below. For more information, refer to Appendix 1.) 6.
Page 72 Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Double stain of green fluorescence dye (Alexa 488) and red fluorescence dye (Alexa 546) Sequential scan (Line Sequential is introduced here.) 1.
Page 73: Double Stain On Xyz Image
Image Acquisition Image Acquisition (Double Stain on XYZ Image) (Double Stain on XYZ Image) Acquisition of 3D images (XYZ) (fluorescence image only) Sample: Double stain of green fluorescence dye (FITC) and red fluorescence dye (Rhodamine) This is the procedure to acquire images through Line Sequential scanning.
Page 74 Image Acquisition Image Acquisition (Double Stain on XYZ Image) (Double Stain on XYZ Image) 9. Select AutoHV and then select ScanSpeed. 10. Select Depth. 11. Press the XYZ button to acquire an image. 12. Click on “SeriesDone”, and “2D View- LiveImage(x)”...
Page 75: Four Stain On Xy Image
Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) Acquisition of 4 stain images (XY) (fluorescence image only) Sample: Four stain of Blue fluorescence dye (DAPI) ,green fluorescence dye (Alexa488) and red fluorescence dye (Rhodamine), far-red fluorescence dye (Cy5) This is the procedure to acquire images through Virtual Channel scan Select Virtual channel...
Page 76 Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) 4. Select “Phase1”, “DAPI”, Alexa488, RhodaminRed are registered on ImageAquisitionControl. * Slit and Filter, DM are automatically set for “DAPI”, “Alexa488” or “PhodaminRed” 5. Select “Phase2”, Cy5 is registered on ImageAquisitionControl * Slit and Filter, DM are automatically set for “Cy5”...
Page 77 Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) 7. Click on Virtual Channel Controller to acquire the image. * Be able to start at each Phase. 8. Saving the image: Right-click on the Image Icon displayed on the Data 8.
Page 78: Single Stain + Dic On Xy Image
Image Acquisition Image Acquisition (Single Stain + DIC on XY Image) (Single Stain + DIC on XY Image) Acquisition of a single image (XY plane) (fluorescence image and differential interference contrast image) Sample: Green fluorescence dye (FITC) and differential interference contrast image 1.
Page 79 Image Acquisition Image Acquisition (Single Stain + DIC on XY Image) (Single Stain + DIC on XY Image) 5. Press the “XY Repeat” button to start scanning. 6. Adjust the green (FITC) image and the differential interference contrast image. 7. Press the “Stop button” to stop scanning.
Page 80: Merge The Image Between Fluorescent Xy Image And Dic Image
Merge the images between Merge the images between fluorescent XY image and image fluorescent XY image and image Edit different each files to the same file. This is available for making merge image Between fluorescent image and focused DIC image. 1.
Page 81: Single Stain On Xyzt Image / Zdc Xyzt Image
Image Acquisition Image Acquisition (Single Stain on XYZT Image) (Single Stain on XYZT Image) This is available for the Time series scan experiment. 1. Adjust the image. Refer P17,18 2. Enter interval time to “Interval” Enter interval number to “Num” Example: Acquiring time series scan images every 5minutes for 1hour is below, 3.
Page 82 Image Acquisition Image Acquisition (Single Stain on XYZT Image) (Single Stain on XYZT Image) 1. Adjust the image. * Refer P17,18 2. Insert ZDC unit to left side. 3. Check “EnableZDC AF during Time Series Scan” and click “ZDC setting” . 4.
Page 83: Reload The Image Conditions
Reload the image conditions Reload the image conditions 1. Open the file and click 2. Click 3. The conditions (HV,Offset, CA and so on ) are reloaded .
Page 84: Image Analysis
Overview of the 2D Operation Panel Overview of the 2D Operation Panel Display switching Click on the button to switch the display Frame advance Enlarge 1:1 display Animation Adjust to the Ch1 display window size Advance speed TEXT Ch2 display Various kinds of ROIs formulation...
Page 85: Image Analysis
Image Analysis (Acquire a Projection Images) Image Analysis (Acquire a Projection Images) 1. Click on the button to select 2. To save this image, right-click on the image, select Save Display and save the image with a new name.
Page 86 Image Analysis Image Analysis (Save a Z section Image as 2D file) (Save a Z section Image as 2D file) Save the image in step 3 or 5 6. Click on the button. 7. A 2D View-(file name) image is created.
Page 87 Image Analysis (Inserting the Scale Bar) Image Analysis (Inserting the Scale Bar) 1. Click on the button. 2. While left-clicking the image, drag and drop it at a certain point. Change the size 3. While clicking the right or left handle, move the mouse from side to side.
Page 88 Image Analysis Image Analysis (Rotating a Three dimensional Image) (Rotating a Three dimensional Image) 1. Click on the button for a 2D View-(file name) image. 2. A 3D view is created. 3. Drag the mouse on the image to observe it at a certain angle. Simple animation 4.
Page 89 Image Analysis (Saving an Image) Image Analysis (Saving an Image) Convert each channel of an XY or XYZ image into a TIFF format 1. Right-click on the Image Icon displayed on the Data Manager and select Export. 2. Set Save as type to TIFF. 3.
Page 90 Image Analysis Image Analysis (Rotating a Three dimensional animation) (Rotating a Three dimensional animation) To save a rotation file as an animated image, create three- dimensional images according to the following procedure. For example, try to rotate an image by 180 degrees.
Page 91: Edit The Image Color And Contrast
2D Image Analysis 2D Image Analysis ( Edit the image color and contrast) ( Edit the image color and contrast) 1. Click “LUT” and then LUT table appears below, 2. Edit contrast to drag to left or right side, and another way to 3.
Page 92: The Image Of Z Section
2D Image Analysis 2D Image Analysis ( the image of Z section ) ( the image of Z section ) 1. Click and select again, then Projection image is shown on 2D View after getting XYZ image. 2. Click and select 3.
Page 93: Intensity Profile Of Each Z Sections
2D Image Analysis 2D Image Analysis ( Intensity Profile of each Z sections ) ( Intensity Profile of each Z sections ) 1. Click and then 2D Control Panel is shown below, 2. Click “Profile” and then Intensity Profile of each Z sections is shown on the X and Y axis.
Page 94: Measure
2D Image Analysis 2D Image Analysis ( Measure ) ( Measure ) 1. Enclose interesting regions by ROI Line on interesting positions by ROI 2. Click “measure”. 4. According to click “Measure All ROIs” ,then the information of all ROI is calculated on Region Measurement.
Page 95 2D Image Analysis 2D Image Analysis ( Line Intensity Profile on the 2D image ) ( Line Intensity Profile on the 2D image ) 1. Line on the 2D image by ROI 2. Click “Intensity Profile” 3. “Intensity Profile” on the line is shown as intensity graph .
Page 96: Line Series Analysis / Co-Localization
2D Image Analysis 2D Image Analysis Line Series Analysis Line Series Analysis 1. Line on the 2D image. 2. Click “Line Series Analysis” 3. Intensity of Z position/ time on the line is shown as a graph . 2D Image Analysis 2D Image Analysis ( Co localization )
Page 97 2D image Analysis(Series Analysis TimeLapse) 1. Enclose interesting regions by ROI 2. Click “Series Analysis” 3. “Series Analysis” graph is shown below, Y axis shows intensity, X axis shows time and then be able to see time series reaction each ROIs.
Page 98: Closing The System
Closing the System Closing the System 1. Exit the FV10-ASW software by selecting File/Exit. 2. Exit the Windows. (1) Select Start/Shut Down. (2) On the Shut Down Window, select Shut Down and click on OK. 3. Turn the laser OFF. (Turn the key switch to the OFF position.) 3-1.
Page 99 Laser Conforcal Scanning Microscope FV1000D Spectral Type (Upright Microscope BX61) Operation Manual...
Page 100 Contents Contents System introdution FV1000D Laser DyeList System Preparation Visible Observation Observation of Fluorescence image Observation of Differential Interference Contrast Images Image Acquisition Overview of Operation Panel for Image Acquisition Single Stain on XY Image 9-11 Complement of adjusting the image 12-13 Double Stain on XY Image 14-15...
Page 101 Spectral Type Main Scanner...
Page 102 Dye List (FV1000D Lasers are available below) LD405nm LD440nm LD473nm LD559nm LD635nm Ar458nm Ar488nm Ar515nm HeNe (G) 543nm...
Page 103: System Preparation
System Preparation System Preparation 1. Turn the computer ON. [In case of equipped concentrated power supply, power on it first] 2. Turn the laser ON (Turning the key switch) 2-1. LD559nm ON 2-2. Multi Ar 458nm 488nm 515nm 2-2. HeNe(G) (643nm) ON 3.
Page 104: Observation Of Fluorescence Image
Visual Observation under the Microscope Visual Observation under the Microscope Observation of Fluorescence Image 1. Select an objective lens by using the hand switch Hand switch 2. Select florescent filter cube MEMO Fluorescence filter NIBA: Blue Excitation / Green Fluorescence (Ex.:FITC,EGFP) WIG: Green Excitation / Red Fluorescence (Ex.:Rhodamine, DsRed)
Page 105: Observation Of Differential Interference Contrast Images
Visual Observation under the Microscope Visual Observation under the Microscope Observation of Differential Interference Contrast Images 1. Select the Objective Lens Hand switch 2. Insert the Polarizing Plate in the Light Pass *DIC prism 3. Insert the DIC prism slider in the light pass Click the button Use this knob...
Page 106: Overview Of Operation Panel For Image Acquisition
Overview of Operation Panel for Image Acquisition Overview of Operation Panel for Image Acquisition Scan mode Scan speed Number of pixels Zoom & Pan Laser output adjustment Objective lens Focus Time Interval & Time Number (for acquisition of XYT or XT image) Transmitted light observation (visual observation)
Page 107: Single Stain On Xy Image
Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Single stain of green fluorescence dye (FITC) 1. Click on the FV10-ASW software button to close the fluorescence lamp shutter.
Page 108 Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) 4. Press XY Repeat button click to get image Continuous scan mode 5. Focus to the specimen 6. Adjust the green (FITC) image. Adjust sensitivity of HV and ·...
Page 109 Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) Select AutoHV and then select ScanSpeed. *As the scan speed becomes slower, noise can be removed while maintaining the current brightness. Press the Stop button to stop scanning Click on XY, and "2D View-LiveImage(x)"...
Page 110: Complement Of Adjusting The Image
Complement of adjusting the image Complement of adjusting the image 1. Click “Clip scan” button , and enclose an interesting region’s image on the whole image. 2. pixel setting *The standard pixel is 512 x 512 3. Zoom Setting Press “XY Repeat” to scan and set zoom value. Zoom Reset button Above image is zoomed From 1x to 2 * Scan speed and pixel resolution remain...
Page 111 Complement of adjusting the image Complement of adjusting the image 5. Pan X,Y Be able to move the field of view to set Pan X,Y without stage action 6. Rotation PanX,Y and Rotation reset button Be able to rotate the whole image. Click “Auto”...
Page 112: Double Stain On Xy Image
Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Double stain of green fluorescence dye (Alexa 488) and red fluorescence dye (Alexa 546) Simultaneous scan 1.
Page 113 Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) 4. Press the XY Repeat button to start scanning. 5. Adjust the green (Alexa Fluor 488) image and the red (Alexa Fluor 546) image. (The image adjustment is outlined below. For more information, refer to Appendix 1.) 6.
Page 114 Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Double stain of green fluorescence dye (Alexa 488) and red fluorescence dye (Alexa 546) Sequential scan (Line Sequential is introduced here.) 1.
Page 115: Double Stain On Xyz Image
Image Acquisition Image Acquisition (Double Stain on XYZ Image) (Double Stain on XYZ Image) Acquisition of 3D images (XYZ) (fluorescence image only) Sample: Double stain of green fluorescence dye (FITC) and red fluorescence dye (Rhodamine) This is the procedure to acquire images through Line Sequential scanning.
Page 116 Image Acquisition Image Acquisition (Double Stain on XYZ Image) (Double Stain on XYZ Image) 9. Select AutoHV and then select ScanSpeed. 10. Select Depth. 11. Press the XYZ button to acquire an image. 12. Click on “SeriesDone”, and "2D View-LiveImage(x)" is displayed on the window bar for the image that has been acquired.
Page 117: Four Stain On Xy Image
Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) Acquisition of 4 stain images (XY) (fluorescence image only) Sample: Four stain of Blue fluorescence dye (DAPI) ,green fluorescence dye (Alexa488) and red fluorescence dye (Rhodamine), far-red fluorescence dye (Cy5) This is the procedure to acquire images through Virtual Channel scan Select Virtual channel...
Page 118 Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) 4. Select “Phase1”, “DAPI”, “Alexa488”, “RhodaminRed” are registered on ImageAquisitionControl. * Slit and Filter, DM are automatically set for “DAPI” “Alexa488” “PhodaminRed” 5. Select “Phase2”, Cy5 is registered on ImageAquisitionControl * Slit and Filter, DM are automatically set for “Cy5”...
Page 119 Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) 7. Click on Virtual Channel Controller to acquire the image. * Be able to start at each Phase. 8. Saving the image: Right-click on the Image Icon displayed on the Data 8.
Page 120: Single Stain + Dic On Xy Image
Image Acquisition Image Acquisition (Single Stain + DIC on XY Image) (Single Stain + DIC on XY Image) Acquisition of a single image (XY plane) (fluorescence image and differential interference contrast image) Sample: Green fluorescence dye (FITC) and differential interference contrast image 1.
Page 121 Image Acquisition Image Acquisition (Single Stain + DIC on XY Image) (Single Stain + DIC on XY Image) 5. Press the “XY Repeat” button to start scanning. 6. Adjust the green (FITC) image and the differential interference contrast image. 7. Press the “Stop button” to stop scanning.
Page 122 Merge the images between Merge the images between fluorescent XY image and image fluorescent XY image and image Edit different each files to the same file. This is available for making merge image Between fluorescent image and focused DIC image. 1.
Page 123 Image Acquisition (Spectral Image on XYL Image) Image Acquisition (Spectral Image on XYL Image) Acquisition of a spectral image (XYL) Sample: Double stain of green fluorescence dye (Alexa Fluor 488) and green fluorescence dye (YOYO-1) 1. Click on the FV10-ASW software button to close the fluorescence lamp shutter.
Page 124 Image Acquisition (Spectral Image on XYL Image) Image Acquisition (Spectral Image on XYL Image) 4. Click on the button, and the Spectral Setting window appears. Set the slit width for CHS1 to 20 nm, for example. 6. Press the XY Repeat button to start scanning.
Page 125 Image Acquisition (Spectral Image on XYL Image) Image Acquisition (Spectral Image on XYL Image) 10. Set the range of wavelength to be acquired, the slit width and the step. Start = Start wavelength End = End wavelength Resolution = Slit width StepSize = Step Select AutoHV and then select ScanSpeed.
Page 126 Image Analysis ( Unmixing Image Analysis ( Unmixing I. When each fluorescence dye point is clear I. When each fluorescence dye point is clear From an XYL image where fluorescence dyes with similar fluorescence spectrums are present together, derive the fluorescence spectrum for each fluorescence dye and obtain an unmixed image based on the fluorescence spectrums.
Page 127 Image Analysis ( Unmixing Image Analysis ( Unmixing I. When each fluorescence dye point is clear I. When each fluorescence dye point is clear Sample: single stain of green fluorescence dye (GFP) and auto fluorescence from cell 1. Open the XYL image (GFP + auto fluorescence).
Page 128: Unmixing
Image Analysis ( Unmixing Image Analysis ( Unmixing II. When a control sample is used II. When a control sample is used From an XYL image with a single type of fluorescence dye, derive the fluorescence spectrum of the dye and obtain an unmixed image based on the fluorescence spectrum.
Page 129 Image Analysis ( Unmixing Image Analysis ( Unmixing III. When only the number of types of fluorescence dyes is known III. When only the number of types of fluorescence dyes is known (Blind Unmixing (Blind Unmixing From an XYL image where fluorescence dyes with similar fluorescence spectrums are present together, obtain an unmixed image based on only the number of types of fluorescence dyes.
Page 130: Reload The Image Conditions
Reload the image conditions Reload the image conditions 1. Open the file and click 2. Click 3. The conditions (HV,Offset, CA and so on ) are reloaded .
Page 131: Image Analysis
Overview of the 2D Operation Panel Overview of the 2D Operation Panel Display switching Click on the button to switch the display Frame advance Enlarge 1:1 display Animation Adjust to the Ch1 display window size Advance speed TEXT Ch2 display Various kinds of ROIs formulation...
Page 132: Image Analysis
Image Analysis (Acquire a Projection Images) Image Analysis (Acquire a Projection Images) 1. Click on the button to select 2. To save this image, right-click on the image, select Save Display and save the image with a new name.
Page 133 Image Analysis Image Analysis (Save a Z section Image as 2D file) (Save a Z section Image as 2D file) Save the image in step 3 or 5 6. Click on the button. 7. A 2D View-(file name) image is created.
Page 134 Image Analysis (Inserting the Scale Bar) Image Analysis (Inserting the Scale Bar) 1. Click on the button. 2. While left-clicking the image, drag and drop it at a certain point. Change the size 3. While clicking the right or left handle, move the mouse from side to side.
Page 135 Image Analysis Image Analysis (Rotating a Three dimensional Image) (Rotating a Three dimensional Image) 1. Click on the button for a 2D View-(file name) image. 2. A 3D view is created. 3. Drag the mouse on the image to observe it at a certain angle. Simple animation 4.
Page 136 Image Analysis (Saving an Image) Image Analysis (Saving an Image) Convert each channel of an XY or XYZ image into a TIFF format 1. Right-click on the Image Icon displayed on the Data Manager and select Export. 2. Set Save as type to TIFF. 3.
Page 137 Image Analysis Image Analysis (Rotating a Three dimensional animation) (Rotating a Three dimensional animation) To save a rotation file as an animated image, create three- dimensional images according to the following procedure. For example, try to rotate an image by 180 degrees.
Page 138: Edit The Image Color And Contrast
2D Image Analysis 2D Image Analysis ( Edit the image color and contrast) ( Edit the image color and contrast) 1. Click “LUT” and then LUT table appears below, 2.Edit contrast to drag to left or right side, and another way to edit contrast is entering value on 3.
Page 139: The Image Of Z Section
2D Image Analysis 2D Image Analysis ( the image of Z section ) ( the image of Z section ) 1. Click and select again, then Projection image is shown on 2D View after getting XYZ image. 2. Click and select 3.
Page 140: Intensity Profile Of Each Z Sections
2D Image Analysis 2D Image Analysis ( Intensity Profile of each Z sections ) ( Intensity Profile of each Z sections ) Click and then 2D Control Panel is shown below, 2. Click “Profile” and then Intensity Profile of each Z sections is shown on the X and Y axis.
Page 141: Measure
2D Image Analysis 2D Image Analysis ( Measure ) ( Measure ) 1. Enclose interesting regions by ROI Line on interesting positions by ROI 2. Click “measure”. 4. According to click “Measure All ROIs”, then the information of all ROI is calculated on Region Measurement.
Page 142 2D Image Analysis 2D Image Analysis ( Line Intensity Profile on the 2D image ) ( Line Intensity Profile on the 2D image ) 1. Line on the 2D image by ROI 2. Click “Intensity Profile” 3. “Intensity Profile” on the line is shown as intensity graph .
Page 143 2D Image Analysis 2D Image Analysis Line Series Analysis Line Series Analysis 1. Line on the interesting region. 2. Click “Line Series Analysis” 3. Intensity of Z position/ time on the line is shown as a graph . 2D Image Analysis 2D Image Analysis ( Co localization )
Page 144 Closing the System Closing the System 1. Exit the FV10-ASW software by selecting File/Exit. 2. Exit the Windows. (1) Select Start/Shut Down. (2) On the Shut Down Window, select Shut Down and click on OK. 3. Turn the laser OFF. (Turn the key switch to the OFF position.) 3-1.
Page 145 Laser Conforcal Scanning Microscope FV1000D Filter Type (Upright Microscope BX61) Operation Manual...
Page 146: Line Intensity Profile On The 2D Image / Histogram
Contents Contents System introduction FV1000D Laser DyeList System Preparation Visible Observation Observation of Fluorescence image Observation of Differential Interference Contrast Images Image Acquisition Overview of Operation Panel for Image Acquisition Single Stain on XY Image 9-11 Complement of adjusting the image 12-13 Double Stain on XY Image 14-15...
Page 147 Filter Type Main Scanner...
Page 148 Dye List (FV1000D Lasers are available below) LD405nm LD440nm LD473nm LD559nm LD635nm Ar458nm Ar488nm Ar515nm HeNe(G)543nm...
Page 149: System Preparation
System Preparation System Preparation 1. Turn the computer ON. [In case of equipped concentrated power supply, power on it first 2. Turn the laser ON (Turning the key switch) 2-1. LD559nm ON 2-2. Multi Ar 458nm 488nm 515nm 2-2. HeNe(G)(643nm) ON 3.
Page 150: Observation Of Fluorescence Image
Visual Observation under the Microscope Visual Observation under the Microscope Observation of Fluorescence Image 1. Select an objective lens by using the hand switch Hand switch 2. Select florescent filter cube MEMO Fluorescence filter NIBA: Blue Excitation / Green Fluorescence (Ex.:FITC,EGFP) WIG: Green Excitation / Red Fluorescence (Ex.:Rhodamine, DsRed)
Page 151: Observation Of Differential Interference Contrast Images
Visual Observation under the Microscope Visual Observation under the Microscope Observation of Differential Interference Contrast Images 1. Select the Objective Lens Hand switch 2. Insert the Polarizing Plate in the Light Pass *DIC prism 3. Insert the DIC prism slider in the light pass Click the button Use this knob...
Page 152: Overview Of Operation Panel For Image Acquisition
Overview of Operation Panel for Image Acquisition Overview of Operation Panel for Image Acquisition Scan mode Scan speed Number of pixels Zoom & Pan Laser output adjustment Objective lens Focus Time Interval & Time Number (for acquisition of XYT or XT image) Transmitted light observation (visual observation)
Page 153: Single Stain On Xy Image
Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Single stain of green fluorescence dye (FITC) 1. Click on the FV10-ASW software button to close the fluorescence lamp shutter.
Page 154 Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) 4. Press XY Repeat button click to get image Continuous scan mode 5. Focus to the specimen 6. Adjust the green (FITC) image. Adjust sensitivity of HV and ·...
Page 155 Image Acquisition Image Acquisition (Single Stain on XY Image) (Single Stain on XY Image) Select AutoHV and then select ScanSpeed. *As the scan speed becomes slower, noise can be removed while maintaining the current brightness. Press the Stop button to stop scanning Click on XY, and “2D View-LiveImage(x)”...
Page 156: Complement Of Adjusting The Image
Complement of adjusting the image Complement of adjusting the image 1. Click “Clip scan” button , and enclose a interesting region’s image on the whole image. 2. pixel setting * The standard pixel is 512 x 512 3. Zoom Setting Press “XY Repeat”...
Page 157 Complement of adjusting the image Complement of adjusting the image 5. Pan X,Y Be able to move the field of view to set Pan X,Y without stage action 6. Rotation PanX,Y and Rotation reset button Be able to rotate the whole image. Click “Auto”...
Page 158: Double Stain On Xy Image
Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Double stain of green fluorescence dye (Alexa 488) and red fluorescence dye (Alexa 546) Simultaneous scan 1.
Page 159 Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) 4. Press the XY Repeat button to start scanning. 5. Adjust the green (Alexa Fluor 488) image and the red (Alexa Fluor 546) image. (The image adjustment is outlined below. For more information, refer to Appendix 1.) 6.
Page 160: Sequential Scan Line Sequential
Image Acquisition Image Acquisition (Double Stain on XY Image) (Double Stain on XY Image) Acquisition of a single image (XY plane) (fluorescence image only) Sample: Double stain of green fluorescence dye (Alexa 488) and red fluorescence dye (Alexa 546) Sequential scan (Line Sequential is introduced here.) 1.
Page 161: Double Stain On Xyz Image
Image Acquisition Image Acquisition (Double Stain on XYZ Image) (Double Stain on XYZ Image) Acquisition of 3D images (XYZ) (fluorescence image only) Sample: Double stain of green fluorescence dye (FITC) and red fluorescence dye (Rhodamine) This is the procedure to acquire images through Line Sequential scanning.
Page 162 Image Acquisition Image Acquisition (Double Stain on XYZ Image) (Double Stain on XYZ Image) 9. Select AutoHV and then select ScanSpeed. 10. Select Depth. 11. Press the XYZ button to acquire an image. 12. Click on “SeriesDone”, and “2D View- LiveImage(x)”...
Page 163: Four Stain On Xy Image
Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) Acquisition of 4 stain images (XY) (fluorescence image only) Sample: Four stain of Blue fluorescence dye (DAPI) ,green fluorescence dye (Alexa488) and red fluorescence dye (Rhodamine), far-red fluorescence dye (Cy5) This is the procedure to acquire images through Virtual Channel scan Select Virtual channel...
Page 164 Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) 4. Select “Phase1”, “DAPI”, Alexa488, RhodaminRed are registered on ImageAquisitionControl. * Slit and Filter, DM are automatically set for “DAPI”, “Alexa488”, “PhodaminRed” 5. Select “Phase2”, Cy5 is registered on ImageAquisitionControl * Slit and Filter, DM are automatically set for “Cy5”...
Page 165 Image Acquisition Image Acquisition (Four Stain on XY Image) (Four Stain on XY Image) 7. Click on Virtual Channel Controller to acquire the image. * Be able to start at each Phase. 8. Saving the image: Right-click on the Image Icon displayed on the Data 8.
Page 166: Single Stain + Dic On Xy Image
Image Acquisition Image Acquisition (Single Stain + DIC on XY Image) (Single Stain + DIC on XY Image) Acquisition of a single image (XY plane) (fluorescence image and differential interference contrast image) Sample: Green fluorescence dye (FITC) and differential interference contrast image 1.
Page 167 Image Acquisition Image Acquisition (Single Stain + DIC on XY Image) (Single Stain + DIC on XY Image) 5. Press the “XY Repeat” button to start scanning. 6. Adjust the green (FITC) image and the differential interference contrast image. 7. Press the “Stop button” to stop scanning.
Page 168 Merge the images between Merge the images between fluorescent XY image and image fluorescent XY image and image Edit different each files to the same file. This is available for making merge image Between fluorescent image and focused DIC image. 1.
Page 169: Reload The Image Conditions
Reload the image conditions Reload the image conditions 1. Open the file and click 2. Click 3. The conditions (HV,Offset, CA and so on ) are reloaded .
Page 170: Image Analysis
Overview of the 2D Operation Panel Overview of the 2D Operation Panel Display switching Click on the button to switch the display Frame advance Enlarge 1:1 display Animation Adjust to the Ch1 display window size Advance speed TEXT Ch2 display Various kinds of ROIs formulation...
Page 171: Image Analysis
Image Analysis (Acquire a Projection Images) Image Analysis (Acquire a Projection Images) 1. Click on the button to select 2. To save this image, right-click on the image, select Save Display and save the image with a new name.
Page 172 Image Analysis Image Analysis (Save a Z section Image as 2D file) (Save a Z section Image as 2D file) Save the image in step 3 or 5 6. Click on the button. 7. A 2D View-(file name) image is created.
Page 173 Image Analysis (Inserting the Scale Bar) Image Analysis (Inserting the Scale Bar) 1. Click on the button. 2. While left-clicking the image, drag and drop it at a certain point. Change the size 3. While clicking the right or left handle, move the mouse from side to side.
Page 174 Image Analysis Image Analysis (Rotating a Three dimensional Image) (Rotating a Three dimensional Image) 1. Click on the button for a 2D View-(file name) image. 2. A 3D view is created. 3. Drag the mouse on the image to observe it at a certain angle. Simple animation 4.
Page 175 Image Analysis (Saving an Image) Image Analysis (Saving an Image) Convert each channel of an XY or XYZ image into a TIFF format 1. Right-click on the Image Icon displayed on the Data Manager and select Export. 2. Set Save as type to TIFF. 3.
Page 176 Image Analysis Image Analysis (Rotating a Three dimensional animation) (Rotating a Three dimensional animation) To save a rotation file as an animated image, create three- dimensional images according to the following procedure. For example, try to rotate an image by 180 degrees.
Page 177: Edit The Image Color And Contrast
2D Image Analysis 2D Image Analysis ( Edit the image color and contrast) ( Edit the image color and contrast) 1. Click “LUT” and then LUT table appears below, 2.Edit contrast to drag to left or right side, and another way to 3.
Page 178: The Image Of Z Section
2D Image Analysis 2D Image Analysis ( the image of Z section ) ( the image of Z section ) 1. Click and select again, then Projection image is shown on 2D View after getting XYZ image. 2. Click and select 3.
Page 179: Intensity Profile Of Each Z Sections
2D Image Analysis 2D Image Analysis ( Intensity Profile of each Z sections ) ( Intensity Profile of each Z sections ) 1. Click and then 2D Control Panel is shown below, 2. Click “Profile” and then Intensity Profile of each Z sections is shown on the X and Y axis.
Page 180: Measure
2D Image Analysis 2D Image Analysis ( Measure ) ( Measure ) 1. Enclose interesting regions by ROI Line on interesting positions by ROI 2. Click “measure”. 4. According to click “Measure All ROIs” , then the information of all ROI is calculated on Region Measurement.
Page 181 2D Image Analysis 2D Image Analysis ( Line Intensity Profile on the 2D image ) ( Line Intensity Profile on the 2D image ) 1. Line on the 2D image by ROI 2. Click “Intensity Profile” 3. “Intensity Profile” on the line is shown as intensity graph .
Page 182: Line Series Analysis / Co-Localization
2D Image Analysis 2D Image Analysis Line Series Analysis Line Series Analysis 1. Line on the interesting region. 2. Click “Line Series Analysis” 3. Intensity of Z position/ time on the line is shown as a graph . 2D Image Analysis 2D Image Analysis ( Co localization )
Page 183: Closing The System
Closing the System Closing the System 1. Exit the FV10-ASW software by selecting File/Exit. 2. Exit the Windows. (1) Select Start/Shut Down. (2) On the Shut Down Window, select Shut Down and click on OK. 3. Turn the laser OFF. (Turn the key switch to the OFF position.) 3-1.
Page 186 OLYMPUS CORPORATION Shinjuku Monolith, 3-1, Nishi Shinjuku 2-chome,Shinjuku-ku, Tokyo, Japan OLYMPUS LIFE SCIENCE EUROPA GMBH Postfach 10 49 08, 20034, Hamburg, Germany OLYMPUS AMERICA INC. 3500 Corporate Parkway, P.O. Box 610, Center Valley, PA 18034-0610, U.S.A. OLYMPUS SINGAPORE PTE LTD...

Olympus FLUOVIEW FV1000 User Manual

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