Olympus FLUOVIEW FVMPE-RS User Manual
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Olympus FLUOVIEW FVMPE-RS User Manual

Olympus FLUOVIEW FVMPE-RS User Manual

Multi photon laser scanning microscope
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User's Manual
FLUOVIEW
FVMPE-RS
Multi Photon Laser Scanning Microscope
Notice
Optical Microscope and Accessory
Thank you for your purchase of Olympus microscope at this time.
Retain this manual in an easily accessible place near a system for future reference.
- Research use only -
AX8334

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Summary of Contents for Olympus FLUOVIEW FVMPE-RS

  • Page 1 Multi Photon Laser Scanning Microscope Notice Optical Microscope and Accessory Thank you for your purchase of Olympus microscope at this time. Retain this manual in an easily accessible place near a system for future reference. - Research use only -...
  • Page 3 • Service “Service” means any adjustment or repair performed by Olympus service personnel or technical personnel who are provided the service training according to the service manual provided by Olympus. The performance has influence on the feature of this system, and there is a risk that unintended CLASS 4 laser light is emitted.

  • Page 4: Registered Trademarks

    Registered Trademarks Registered Trademarks Microsoft, Microsoft Windows, Excel for Windows are registered trademarks of Microsoft Corporation. Mai Tai ® , InSigh , and DeepSee are registered trademarks of Newport Corporation. Chameleon , OBIS , and Sapphire are registered trademarks of Cohelent Inc.. ProScan is registered trademark of Prior Scientific Ltd..

  • Page 5: Safety Manual

    Safety Manual On this volume This volume describes warnings and cautions for your safety use of this system. Before use, be sure to observe all the warnings and cautions. In case this system is used in other methods than those described in this manual, the safety cannot be insured and a system failure may occur.
  • Page 7 In accordance with European Directive on Waste Electrical and Electronic Equipment, this symbol indicates that the product must not be disposed of as unsorted municipal waste, but should be collected separately. Refer to your local Olympus distributor in EU for return and/or collection systems available in your country. Research Use Only This product does not conform to FCC Part15 Class A.

  • Page 8: Table Of Contents

    Contents 1 Safety caution ............................ 1-1 Transportation and modification of this system ................1 1-2 Waste of system ................................1 1-3 Handling precautions ..............................2 Mechanical (heat) precaution ..............................2 Electrical precaution ....................................3 Laser precaution .....................................4 1-4 Necessary Actions ..............................8 Appointing a laser safety officer ..............................8 Laser controlled areas (warning signs and off-limits) ....................9 Preparing and wearing preventive eye glasses and protective clothes ..........10 Ophthalmologic examination ..............................10...

  • Page 9: Safety Caution

    • The used mercury lamp in this system must be disposed of as industrial waste. If you cannot dispose CAUTION of it properly, please contact Olympus. • In order to replace the lithium coin battery built in the touch panel controller in this system, please contact our sales office in your local area.

  • Page 10: Handling Precautions

    1-3 Handling precautions CAUTION • Also, refer to the part related to safety in manuals of surrounding units. However, if the following units are used in this system, the manuals provided with those units are not effective. See this manual about usage of those units. •...

  • Page 11: Electrical Precaution

    EMC performance of the device can not be assured. • Always use the power cord provided by Olympus. If no power cord is provided, please select the proper power cord by referring to the section "PROPER SELECTION OF THE POWER CORD" at the end of the safety manual.

  • Page 12: Laser Precaution

    Laser precaution This system is specified as a laser product of the class as shown below, depending upon the laser to be combined. (1) Laser combination CLASS 4 (IEC60825-1:2007) This product complies with 21 CFR 1040.10 and 1040.11 except for deviations pursuant to Laser Notice No.50, dated June 24, 2007 Laser products of Class 4 are products that can only be used under control of laser safety officer.
  • Page 13 FVMPE-RS • Direct light, specular reflected light and diffused reflection light of laser are dangerous. CAUTION • The laser used in this system is dangerous because the wavelength of the laser is an invisible near- infrared light. • Total laser radiation emitted from this system are as follows: MPE (Maximum Permissible Exposure): 0.016 J/m (for a single pulse)
  • Page 14 • Do not bend or pull the optical fiber cable with an excessive force or step on it. If the optical fiber cable is damaged, the laser light may leak outside it and cause an extremely hazardous situation. Should such an event occur, immediately turn off the laser power and contact your local Olympus representative.
  • Page 15 FVMPE-RS Eye damage CAUTION • Invisible laser emitted from this system focuses on retina of the eyeball. • If the laser enters into your eyes, the retina may be damaged to result in blindness, etc. • Get the protective eyewear matching class and wavelength of the laser of this system. •...

  • Page 16: Necessary Actions

    1-4 Necessary Actions OLYMPUS would like to take this opportunity to ask the customer to establish a system to ensure laser safety in line with the customer’s facility. The items necessary for ensuring laser safety are listed below. Appointing a laser safety officer Please take actions to ensure laser safety with a laser safety officer at the center of such activities.

  • Page 17: Laser Controlled Areas (Warning Signs And Off-Limits)

    FVMPE-RS Laser controlled areas (warning signs and off-limits) The area where a laser beam of class 3B or above is emitted is defined as laser control area. Define the room where this system is set up as laser control area. Place a warning sign near to each of the laser controlled areas that includes the following information: ·...

  • Page 18: Preparing And Wearing Preventive Eye Glasses And Protective Clothes

    Preparing and wearing preventive eye glasses and protective clothes In Laser Hazard Area, it is essentially required to wear eye protector and protective clothing. For this reason, in case a microscope should be operated, or a specimen should be set, wear the eye protector. 1 Using protective glasses Get the protective glasses matching class and wavelength of the laser of this system.

  • Page 19: Safety Functions Of This System

    This system is provided with the safety measures corresponding to the specifications required for class 4 laser products. CAUTION These safety functions are ensured in the state in which OLYMPUS delivers the microscope. Do not attempt to disassemble the microscope or the laser unit.

  • Page 20: Warning Labels

    Warning labels Safety symbols The following symbols are found on this system. Study the meaning of the symbols, and always use this system in the safest possible manner. Symbol Explanation Indicates that the surface becomes hot, and should not be touched with bare hands.

  • Page 21: Laser Safety Warning Labels

    FVMPE-RS 2-1 Laser safety warning labels Warning labels Type of labels attached to a laser system may be different, depending upon laser combination in your possession. Location attached • FV30-HSUMP Hybrid scanning unit for MPE (FV30-HSUMP)

  • Page 22: Aperture Label

    Aperture label Laser emits from the place where the above label exists and the arrow indicates. • BX63LF Laser aperture External laser...
  • Page 23 FVMPE-RS • FV30GF Laser aperture External laser...

  • Page 24: Protective Housing Labels

    Protective housing labels } As labels are attached to many places, the label positions are shown per unit later in this section. Product unit name BX63LF FV30-COMB4 FV30-LCU FV30-LCUMP 405nm Laser FV30- ADP405 Non descan DM 458nm Laser FV30- ADP458 FV30-NDM690 LSM Mirror FV30-NDM760...
  • Page 25 FVMPE-RS FV30GF FV30-COMB4 FV30-LCU FV30-LCUMP 405nm Laser FV30- ADP405 458nm Laser FV30- ADP458 FV30- U-AW 588nm Laser FV30-LCUMP U-FFWADP ADP588 U-LLG300 U-HGLGPS Non descan DM FV30-NDM690 LSM Mirror FV30-FURBB FV30-NDM760 Fluorescence Cube DM for SIM scanner FV30-NDMVCOIR FV30-ADM800 U-IFCBL100 FV30-LASA FV30-LAUS FV30-SIMMP FV30-ADMOPT...
  • Page 26 One laser system IR Laser FV30-LASA FV30-LAUS FV30-LASB IR Laser FV30-LASA FV30-LAUS FV30-LASB...
  • Page 27 FVMPE-RS Twin laser system IR Laser IR Laser FV30-LAUT FV30-LATAA FV30-LATAB IR Laser IR Laser FV30-LATAA FV30-LATAB FV30-LAUT...
  • Page 28 Dual lines system IR Laser FV30-LADAB FV30-LAUT IR Laser FV30-LADAB FV30-LAUT...
  • Page 29 FVMPE-RS Microscope and scan unit BX63LF FV30-HSUMP FV30-ETTR FV30-TBSUB FV30-RFABXL U-LLG300 FV30-TXDBX FV30-LAUS BX63L-NPA40 BX63LF BX63L-SVLAD FV30-BPBXL FV30GF FV30-ETTR FV30-HSUMP FV30-TBSUG FV30-RFABXL U-LLG300 FV30-LAUS FV30GF...
  • Page 30 BX63LF FV30-SIMMP FV30-HSUMP FV30-ETTR FV30-TBSIMB FV30-FURBB FV30-RFABXL FV30-TBSUB U-LLG300 FV30-TXDBX FV30-LAUT BX63L-NPA40 BX63LF BX63L-SVLAD FV30-BPBXL FV30GF FV30-ETTR FV30-SIMMP FV30-HSUMP FV30-TBSIMG FV30-TBSUG FV30-RFABXL FV30-FURBB U-LLG300 FV30-LAUT FV30GF...
  • Page 31 FVMPE-RS Detector FV30-RXD FV30-TXDBX FV30-ARXD FV30-RXD FV30-TXDBX FV30-AGAPD FV30-RXD FV30-TXDBX...
  • Page 32 Detail view • FV30-HSUMP...
  • Page 33 FVMPE-RS • FV30-SIMMP...
  • Page 34 • FV30-LAUS One laser system • FV30-LASA FV30-LASB...
  • Page 35 FVMPE-RS • FV30-LATAA FV30-LATAB Twin laser system • FV30-LAUT Dual lines system • FV30-LADAB...
  • Page 36 • FV30-TBSUB FV30-TBSIMB...
  • Page 37 FVMPE-RS • FV30-TBSUG FV30-TBSIMG...
  • Page 38 • FV30-RFABXL BX63LF...
  • Page 39 FVMPE-RS • FV30-RFABXL FV30GF...
  • Page 40 • FV30-RXD Open...
  • Page 41 FVMPE-RS • FV30-ARXD Open...
  • Page 42 • FV30-AGAPD Open...
  • Page 43 FVMPE-RS • FV30-TXDBX...
  • Page 44 • FV30-TD...
  • Page 45 FVMPE-RS • FV30-COMB4 • FV30-ADP405 • FV30-ADP458 • FV30-ADP588...
  • Page 46 • FV30-FURBB...

  • Page 47: Heat Safety Warning Label

    FVMPE-RS 2-2 Heat safety warning label • U-LH100IR...
  • Page 48 • U-LH100...
  • Page 49 FVMPE-RS • U-AW...
  • Page 50 • U-HGLGPS...

  • Page 51: Finger Safety Warning Label

    FVMPE-RS 2-3 Finger safety warning label...
  • Page 52 • BX63LF Though protection bin labels are attached to other places than the places indicated in this manual, the said labels are intended for our service personnel. CAUTION In case the warning label is blurred or peeled off and, if it should be replaced or if you need help, contact our local sales office immediately.

  • Page 53: Specifications

    FVMPE-RS Specifications 3-1 Specifications of units · Dimensions and weights Unit Specifications FV30-COMB4 Dimensions (mm) 400 (Depth) X 402 (Width) X 200 (Height) Weight 12.4 kg BX63LF Dimensions (mm) Microscope frame: 294.4 (Depth) X 413.2 (Width) X 347.5 (Height) Touch panel controller: 211 (Depth) X 190 (Width) X 105 (Height) Weight Microscope frame: 11 kg Touch panel controller: 2.4 kg...
  • Page 54 Unit Specifications BX63L-SVLAD Dimensions (mm) Long pillar unit: Ø 16 X 200.4 (Height) Middle pillar unit: Ø 16 X 160.4( (Height) Short pillar unit: Ø 16 X 46 (Height) Weight Long pillar unit: 0.3 kg Middle pillar unit: 0.23 kg Short pillar unit: 0.04 kg XLPLN25XWMP2 Dimensions (mm)
  • Page 55 FVMPE-RS Unit Specifications FV30-TXDBX Dimensions (mm) 333 (Depth) X 288 (Width) X 133 (Height) Weight 5.2 kg FV30-LCU Dimensions (mm) 510 (Depth) X 426 (Width) X 194 (Height) Weight 14 kg FV30-FURBB Dimensions (mm) Coupling lens unit: Ø 24 X 63.5 (Length) Fiber unit: Ø...
  • Page 56 Unit Specifications FV30-LAUS Dimensions (mm) 580 (Depth) X 565 (Width) X 180 (Height) Weight 15.5 kg FV30-LAUT Dimensions (mm) 750 (Depth) X 635 (Width) X 180 (Height) Weight 30.5 kg FV30-LCUMP Dimensions (mm) 510 (Depth) X 426 (Width) X 195 (Height) Weight 13.6 kg FV30-LASA...
  • Page 57 FVMPE-RS · FVMPE-RS Power supply rating Item Name Rating FV30-PSU Power supply and electronic control unit AC 100-120 V/220-240 V 5.0 A/2.3 A 50/60 Hz FV30-LCU Laser control unit AC 100-120 V/220-240 V 1.2 A/0.63 A 50/60 Hz FV30-LCUMP MPE laser control unit AC 100-120 V/220-240 V 1.5 A/0.76 A 50/60 Hz...

  • Page 58: Specifications Of Compatible Lasers

    690-1040 nm Chameleon VisionII Olympus 680-1080 nm Chameleon Vision S Olympus 690-1050 nm } For detail specifications of the IR pulse laser, refer to the materials provided by COHERENT or Spectra-Physics. } There are items which must be confirmed before starting or exiting the IR pulse laser. For details, refer to "2-5...
  • Page 59 FVMPE-RS } The table below shows the tolerance of the laser entering each port. FV30-HSUMP FV30-SIMMP IR port IR port Vis port Applicable 700-1300 nm 700-1100 nm 400-650 nm wavelength Tolerable 764 W/cm 382 W/cm 6.4 W/cm power density Converted to average power Converted to average power ·...

  • Page 60: Proper Selection Of The Power Supply Cord

    If no power supply cord is provided, please select the proper power supply cord for the equipment by referring to “Specifications” and “Certified Cord” below; CAUTION: In case you use a non-approved power supply cord for Olympus products, Olympus can no longer warrant the electrical safety of the equipment.
  • Page 61 FVMPE-RS Table 2 HAR flexible cord Approval organizations and cordage harmonization marking methods Alternative Marking Utilizing Printed or Embossed Harmoniza- Black-Red-Yellow Thread tion Marking (May be located on (Length of color section in mm) Approval Organization jacket or insulation of internal wiring) Black Yellow...
  • Page 63 Operation / Maintenance Manual...
  • Page 65 FVMPE-RS Precautions and notes for glare prevention When excitation filter is removed from the fluorescence mirror unit When the excitation filter is removed from the fluorescence mirror unit and attached on the excitation filter slider or exci- tation filter wheel on the side of the white lamp, very glaring light (*1) may enter the eyepieces in the following cases. When the illumination light from the white light lamp (*2) is input without passing through an excitation filter.
  • Page 66 Opening and closing of the shutter to shield the light of the white light lamp By pushing the FL SHUTTER button on the attached controller, U-MCZ, you can switch between irradiation and blocking of the illumination light. FL SHUTTER Button* * The FL SHUTTER button opens/closes the reflected shutter only.
  • Page 67 Contents 1 System outline ..................................1 1-1 Features of FVMPE-RS ....................................1 1-2 Features of each system ..................................2 External NDD (Non-Descanned Detector) unit ..............................2 1-3 Allowable work area when attaching/removing units...................3 1-4 System configuration ....................................5 System diagram ..............................................5 1-5 Confirmation when working with FVMPE-RS .........................7 Key operation ................................................7 Checking the emission indicator .....................................7 2 Precautions for use ...............................8...
  • Page 68 4-2 Stages (Operation/Maintenance)..............................18 How to install the stage (Maintenance) ..................................18 Placing the specimen (Operation) ....................................21 Moving the specimen (Operation) ....................................22 Setting the grounding (Maintenance) ..................................23 Adjusting the X-axis/Y-axis knob rotation tension (Operation) ....................................... . 23 Using the light shielding sheet (Operation)...............................24 4-3 Microscope BX63LF (Operation/Maintenance) ......................25 Using the Filters (Maintenance) (BX63LF only) ..............................25...
  • Page 69 4-7 Mercury lamp (for U-HGLGPS) (Operation) ........................35 Turning on the lamp ............................................35 Turning off the lamp ............................................35 Adjustment of the brightness ......................................36 Counter display ..............................................36 Lighting display LED ............................................36 5 How to assemble each NDD unit ......................38 5-1 External NDD unit (reflected right) (Maintenance)....................39 Unit structures ................................................39 Attaching and removing the cover (FV30-RXD) ..............................41 Attaching and removing the fluorescence cube (FV30-RXD) .......................42...
  • Page 70 5-4 Fluorescent mirror unit and DM unit (Maintenance) ....................56 Fluorescence mirror unit (IX3-FFXL)....................................56 DM Unit for the SIM Scanner (FV30-ADMOPT) ...............................59 6 Visual observation / observation using a camera ..............60 6-1 Touch panel controller (Operation/Maintenance) ....................60 Basic operations of touch panel controller (Operation/Maintenance) ..................60 Starting the touch panel controller (Operation/Maintenance) .
  • Page 71 Differential interference contrast (DIC) observation (Operation/Maintenance) 72 Adjusting the polarizer position (Maintenance) ..............................72 Observation method (Operation).......................................75 7 LSM observation (Operation) ..........................76 8 Signal connection with external device ....................77 8-1 Features of FV30-ANALOG ..................................77 8-2 Specifications of FV30-ANALOG ..............................77 8-3 Setting procedure ......................................81 Setting analog input range from external devices .............................81 Connection ...................................................82 Acquiring analog voltage input signal ..................................82...

  • Page 73: System Outline

    FVMPE-RS System outline 1-1 Features of FVMPE-RS · It is possible to perform high-speed observation using the resonant scanner. · High definition can be achieved using the galvanometer scanner. · It is possible to generate automatically a beam most suitable to the objective lens and laser wavelength, providing high efficiency and resolution imaging.

  • Page 74: Features Of Each System

    1-2 Features of each system External NDD (Non-Descanned Detector) unit The following four types of units are available depending on the combination of detection channels: Number of detection Combination units Application channels FV30-RXD Multi-photon-excited reflected fluorescence observation FV30-RXD, FV30-ARXD Multi-photon-excited fluorescence observation FV30-RXD, FV30-AGAPD Multi-photon-excited fluorescence high-sensitivity observation FV30-TD...

  • Page 75: Allowable Work Area When Attaching/Removing Units

    CAUTION stop radiating the laser before working in this area. • Olympus service personnel, or technical personnel who received service training, will carry out with the attachment to and removal segmentated as service, in accordance with the FVMPE-RS Service Manual.
  • Page 76 CAUTION stop radiating the laser before working in this area. • Olympus service personnel, or technical personnel who received service training, will carry out with the attachment to and removal segmentated as service, in accordance with the FVMPE-RS Service Manual.
  • Page 77 FVMPE-RS...

  • Page 79: Confirmation When Working With Fvmpe-Rs

    • If some lamps are turned ON and some lamps are turned OFF among four lamps, stop using the system and contact Olympus. • Do not remove the cable of the lamp box or hide the lamp box by covering the lamp box, etc. Doing so falls under the remodeling, and is not in conformance with Laser Safety Standards.

  • Page 80: Precautions For Use

    Precautions for use • The IR pulse laser used for multiphoton excitation is a high-power invisible laser that corresponds to CAUTION CLASS 4, making even its scattered light dangerous. Care is required when handling this system, even though it is structurally designed so the power of emitted light is equivalent to CLASS 1 or lower, except light emitted from the objective lens.
  • Page 81 (PC) data, resulting from use of or inability to use this product. • The controller (PC) used with this system has already been set up with Microsoft Windows. This operating system should be backed up and the backup retained carefully by the user. (Olympus does not provide backup support.) For details on the controller (PC) and Microsoft Windows, also refer to the manuals provided with them.

  • Page 82: Turning On/Off The Power And Starting/Exiting From The Software

    Turning on/off the power and starting/exiting from the software 3-1 Turning the power on (Operation) } Be sure to follow the order to turn ON the power described in Step 2 and Step 3. 1. Set the power switches of the controller (PC) and the monitor to I (ON). 2.
  • Page 83 6. Logon windows with your own user ID. } If you have not yet registered your user ID, logon with the factory default ID. Factory default ID : olympus Password : olympus Be sure to keep running the powers of following units of the IR pulse laser around the clock.

  • Page 84: Starting The Software (Operation)

    3-2 Starting the software (Operation) • Turn ON the microscope and power supply units. If the laser control software made by Spectra-Physics CAUTION has been started, exit it before starting this software. • The external NDD unit has extremely high light-receiving sensitivity; therefore, it may deteriorate if it subjected to ambient light from room illumination or a monitor.

  • Page 85: Completing The Observation With This System (Operation)

    FVMPE-RS 3-3 Completing the observation with this system (Operation) 1. Uncheck the checkbox of the laser on the FV30S-SW software to stop emitting the laser first, and exit the FV30S-SW software. } Before exiting the software, you can select ON/OFF (stop oscillation) of the emission of the IR Pulse laser in the [Configuration] dialog box selected in the [Tools] menu.

  • Page 86: Actions Before The

    "Objective Table" using Hand Held Controller. by Spectra-Physics and confirm that ”100XA0.01OL2” is set on the Objective screen. (Note1) Olympus has confirmed the descriptions in the materials after following versions. MAITAI DS Series : 90038200Rev.B Insight DS Series : 90044047Rev.A Chameleon Vision Series : 1086757Rev.AD.

  • Page 87: How To Operate Each Part

    FVMPE-RS How to operate each part From "4-1 Objective lens (Operation/Maintenance)" to "4-5 Observation tube (Operation)" in this chap- CAUTION ter, the maintenance works are included in necessary laser opening area during LSM observation. Be sure to stop radiating the laser before starting relevant maintenance works. 4-1 Objective lens (Operation/Maintenance) •...

  • Page 88: Adjusting The Objectives (Bx63L-Dsgnps Only) (Maintenance)

    Adjusting the objectives (BX63L-DSGNPS only) (Maintenance) } Perform the parfocal adjustment and the centering adjustment of two objectives to be used. Parfocal adjustment procedures Check the difference in Z positions when bringing the specimen into focus with the objective on the front side and when with the objective on the back side by using the touch panel controller.

  • Page 89: Using Immersion Objectives (Operation)

    FVMPE-RS Using immersion objectives (Operation) } Using the immersion objectives to immerse the tip of the objectives in a medium of the specimens allows you to observe the cultured tissue specimens which are often very think. } Recommended immersion objectives: •...

  • Page 90: Stages (Operation/Maintenance)

    4-2 Stages (Operation/Maintenance) Be sure to confirm the laser emission status before starting to work. Refer to page 7 for details. Working CAUTION with the stage area is dangerous due to the opening area of the laser. • If you want to use other stage holders than those provided with this system, be sure not to use the stage holders which reflect the light or are flammable.
  • Page 91 FVMPE-RS FV30GF • When carrying the stage, be sure to hold the knob @ CAUTION (upper picture on next page) of the gate frame microscope exclusive removable plate (FV30-GFRMP) by two people. • When attaching or removing the stage, rotate the Z transfer knob ²...
  • Page 92 } The picture shows that following three units are configured on the removable plate for gantry frame (FV30-GFRMP). Plain stage for gantry frame (FV30-GFSP) Z stage for gantry frame (FV30-GFZST) XY stage for gantry frame (FV30-GFXYST) } Even though the different stage is configured on the gate frame microscope exclusive removable plate (FV30-GFRMP), the same procedures apply when attaching or removing the stage.

  • Page 93: Placing The Specimen (Operation)

    FVMPE-RS Placing the specimen (Operation) BX63LF Place the specimen on the center of the stage. } The optional stage center plate (IX-CP50) makes it possible to observe a wide range of a big petri dish, etc. (Central hole diameter: 50 mm) FV30GF Place the specimen on the center of the stage.

  • Page 94: Moving The Specimen (Operation)

    Moving the specimen (Operation) BX63LF The specimen can be moved by turning the X-axis knob Y-axis knob The movement strokes are 50 mm (X-axis) x 43 mm (Y-axis). FV30GF Loosen the screws @ placed at 2 positions on the XY stage for gantry frame (FV30-GFXYST).

  • Page 95: Setting The Grounding (Maintenance)

    FVMPE-RS Setting the grounding (Maintenance) BX63LF only } In case of electrical physiological experiment, etc., the specimen can be grounded from the stage. } Set the grounding together with setting the grounding of the microscope BX63LF described on page 25. Prepare a grounding wire and M4 screw and attach grounding.

  • Page 96: Using The Light Shielding Sheet (Operation)

    Using the light shielding sheet (Operation) BX63LF only } During fluorescence observation using a low-magnification objec- tive, the fluorescence image may be deteriorated due to light reflected from the condenser or the surroundings. In this case, use the light shielding sheet. Lower the condenser to the lower limit position using the condenser height adjustment knob.

  • Page 97: Microscope Bx63Lf (Operation/Maintenance)

    FVMPE-RS 4-3 Microscope BX63LF (Operation/Maintenance) Be sure to confirm the laser emission status before starting to work. Refer to page 7 for details. Working CAUTION with the microscope (BX63LF) is dangerous due to the opening area of the laser. Using the Filters (Maintenance) (BX63LF only) A filter with a diameter of 45 mm can be inserted in the filter turret attached to the microscope BX63LF base.

  • Page 98: Condenser (Operation/Maintenance)

    4-4 Condenser (Operation/Maintenance) Be sure to confirm the laser emission status before starting to work. Refer to page 7 for details. Working CAUTION with the condenser is dangerous due to the opening area of the laser. Mounting and removing the condenser (Maintenance) } Same procedures apply when mounting or removing either the condenser provided with FV30-TXDBX, WI-UCD or WI-DICD.

  • Page 99: With The Wi-Ucd Condenser (Maintenance)

    FVMPE-RS With the WI-UCD condenser (Maintenance) } There are three types of compatible condensers: WI-UCD, WI-DICD and condenser provided with FV30-TXDBX. List of DIC System Combinations DIC prism for condenser DIC prism for nosepiece Objectives Condenser provided with FV30-TXDBX WI-UCD Oil top lens Dry top lens WI-DICD...
  • Page 100 With the WI-UCD condenser } When selecting the bright field (BF) light path using the WI-UCD, leave one DIC prism (large) mount position empty. Remove the WI-UCD condenser from the microscope frame. Remove the condenser cover by loosening the retaining screws using a coin, etc.

  • Page 101: Centering The Condenser (Operation)

    FVMPE-RS Centering the condenser (Operation) Set the specimen onto the stage. Set the observation optical path of the microscope to [BI 100% (eyepiece 100%)]. Set the fluorescent mirror unit cassette position to [Blank] Open the aperture diaphragm. Adjust the illumination brightness by pushing the two transmitted light brightness control buttons Focus on the specimen.

  • Page 102: Using The Aperture Iris Diaphragm (Operation)

    12. Gradually open the field iris diaphragm. The condenser is properly centered if the iris image is centered and inscribed in the field of view. Before starting the LSM observation, be sure to open the field CAUTION diaphragm and the aperture diaphragm completely. Using the aperture iris diaphragm (Operation) } In general, the potential resolving power of an objective is fully 70-80%...

  • Page 103: Observation Tube (Operation)

    FVMPE-RS 4-5 Observation tube (Operation) Adjusting the interpupillary distance While looking through the eyepieces, adjust the binocular vision until the left and right fields of view coincide completely. The index dot · indicates the interpupillary distance. } Note your interpupillary distance so that it can be quickly duplicated. Adjusting the diopter } The diopter adjustment makes it possible to reduce the specimen focusing error even after the objective is switched.

  • Page 104: Using The Eye Shades

    Using the eye shades When wearing eyeglasses Use the eye shades in the normal, folded-down position. This will prevent the eyeglasses from being scratched. When not wearing eyeglasses Extend the folded eye shades in the direction of the arrow to prevent extraneous light from entering between the eyepieces and eyes.

  • Page 105: Mounting The Camera (Maintenance)

    FVMPE-RS 4-6 Mounting the camera (Maintenance) Mounting a camera } Use the port of Erect image tilting trinocular (FV30-ETTR) when shooting a video or digital camera picture of the microscope images. The trinocular tube accepts a camera adapter (certain camera adapters necessitate a camera mount adapter).
  • Page 106 2 Selecting the camera adapter magnification The image pickup area is determined by the size of the image sensor used in the TV camera or digital camera and the magnification of the camera adapter. The following illustrations show the image pickup areas of camera adapters when the WHN10X eyepieces with a FN of 22 are used.

  • Page 107: Mercury Lamp (For U-Hglgps) (Operation)

    FVMPE-RS 4-7 Mercury lamp (for U-HGLGPS) (Operation) Turning on the lamp Turn “ I ” (ON) the power switch on the back side of the light source unit. The software version appears in the counter, and then (after approximately 3 seconds), the accumulated lighting time of the lamp appears.

  • Page 108: Adjustment Of The Brightness

    Adjustment of the brightness Rotate the light volume adjustment dial to adjust the light volume. The light volume can be adjusted in 7 levels. Each numerical number indicates the percentage (0—100%) of each light volume where the maximum light volume is 100%. } Rotate the light volume adjustment dial slowly by each level.
  • Page 109 Does the lamp lighting time Replace the lamp. exceed 2,000 hours? Trained Olympus service or technical personnel will replace the lamp in accordance with the FVMPE-RS Service Manual. Turn (ON) the power switch, and then press the lamp switch.

  • Page 110: How To Assemble Each Ndd Unit

    At this time, the GaAsP- PMT must be replaced with a new one (paid) or adjusted by the Olympus service personnel. This operation, replacement and adjustment, is paid for, even if it will happen within a year from the delivery.

  • Page 111: External Ndd Unit (Reflected Right) (Maintenance)

    FVMPE-RS 5-1 External NDD unit (reflected right) (Maintenance) Unit structures For 2-channel For 4-channel For 4-channel (FV30-RXD) (FV30-RXD, FV30-ARXD) (FV30-RXD, FV30-AGAPD) IN/OUT selector knob for IN/OUT selector knob for Spectral DM or (N/A) Spectral DM Mirror IR cut filter**** IR cut filter**** IR cut filter**** (N/A) Spectral DM**...
  • Page 112 FV30-ARXD or FV30-AGAPD Š FV30-RXD...

  • Page 113: Attaching And Removing The Cover (Fv30-Rxd)

    FVMPE-RS Attaching and removing the cover (FV30-RXD) 1. Remove the cover of the external NDD unit by loosening and removing the fixing screws (five locations) securing the cover. Cover fixing screws 2. After attaching or removing the part, reposition the cover and secure it by tightening the cover fixing screws. When the cover is removed, the interlock switch works to block the laser beam.

  • Page 114: Attaching And Removing The Fluorescence Cube (Fv30-Rxd)

    Attaching and removing the fluorescence cube (FV30-RXD) 1. Loosen the screws securing the fluorescence cube mount dovetail on the external NDD unit using the Allen screw- driver provided with the system, and remove the fluorescence cube by pulling it out. 2.

  • Page 115: Attaching And Removing The Spectral Dm Cube (Fv30-Rxd)

    FVMPE-RS Attaching and removing the spectral DM cube (FV30-RXD) } When attaching/removing the spectral DM cube, pull the IN/OUT selector knob and hold it to prevent the DM secur- ing table from moving. 1. Loosen the screws securing the spectral DM cube mount dovetail on the external NDD unit using the Allen screw- driver provided with the system, and remove the spectral DM cube by pulling it out.

  • Page 116: Attaching And Removing The Ir Cut Filter (Fv30-Rxd)

    Attaching and removing the IR cut filter (FV30-RXD) } The IR cut filter (BA685RXD) is installed on delivery. 1. Loosen the screw (one location) securing the IR cut filter on the external NDD unit using the Allen screwdriver provided with the system, and remove the IR cut filter. IR cut filter IR cut filter fixing screw 2.

  • Page 117: Attaching And Removing The Cover (Fv30-Arxd, Fv30-Agapd)

    FVMPE-RS Attaching and removing the cover (FV30-ARXD, FV30-AGAPD) 1. Remove the cover of the external NDD unit by loosening and removing the fixing screws (four locations) securing the cover. Projection inside the cover Cover fixing screws } The picture above shows the attaching/removing of the cover for FV30-ARXD. ( Same procedures apply to FV30-AGAPD when attaching/removing the cover.

  • Page 118: Attaching And Removing The Fluorescence Cube (Fv30-Arxd, Fv30-Agapd)

    Attaching and removing the fluorescence cube (FV30-ARXD, FV30-AGAPD) 1. Loosen the screws securing the fluorescence cube mount dovetail on the external NDD unit using the Allen screw- driver provided with the system, and remove the fluorescence cube by pulling it out. 2.

  • Page 119: External Ndd Unit (Transmitted Light) (Maintenance) (Bx63Lf Only)

    FVMPE-RS External NDD unit (Transmitted light) (Maintenance) (BX63LF only) Unit structures (FV30-TXDBX) For 2-channel (FV30-TXDBX) IR cut filters** Fluorescence cubes* PMT channel 1 [TXD1] PMT channel 2 [TXD2] The following fluorescence cubes can be mounted/removed. FV30-FVG, FV30-FCY, FV30-F475R, FV30-FGR, FV30-FRCY5, FV30-FOPT The following IR cut filters can be mounted/removed.

  • Page 120: Removing And Installing The Water Separator (Fv30-Txdbx)

    Removing and installing the water separator (FV30-TXDBX) 1. Pull the water separator secured to the top surface of the external NDD unit with the two holders horizontally to the left in order to remove it. To reinstall the water separator, align the two U-shaped grooves with the two holders on the top surface of the external NDD unit, and then push the water separator down.

  • Page 121: Removing And Installing The Fluorescence Cube (Fv30-Txdbx)

    FVMPE-RS Removing and installing the fluorescence cube (FV30-TXDBX) } The following fluorescence cubes can be mounted/removed. FV30-FVG, FV30-FCY, FV30-F475R, FV30-FGR, FV30-FRCY5, FV30-FOPT 1. Hold the knob of the fluorescence cube and remove the fluorescence cube by pulling it out from the NDD unit. 2.

  • Page 122: Removing And Installing The Ir-Cut Filter (Fv30-Txdbx)

    Removing and installing the IR-cut filter (FV30-TXDBX) } The following IR cut filters can be mounted/removed. FV30-BA750TXD, BA685TXD (installed to FV30-TXDBX on delivery.) 1. Loosen the screw (one location) securing the IR cut filter on the external NDD unit using the Allen screwdriver provided with the system, and remove the IR cut filter.

  • Page 123: Assembling The Fluorescence Cube And The Spectral Dm Cube (Maintenance)

    FVMPE-RS Assembling the fluorescence cube and the spectral DM cube (Maintenance) It is possible to create a spectral mirror unit with wavelength characteristics, according to the needs of the customer by using a commercially available dichroic mirror and emission filter. Unit name External NDD units used Fluorescence cube...
  • Page 124 } Be careful not to soil the dichroic mirror surface with sebum, etc. or scratch it with tools, etc. } Installing the dichroic mirror in the wrong direction will adversely affect optical performance. For Olympus dichroic mirrors, a corner of the interference film surface is marked with a marker. For other commercially...
  • Page 125 } Installing the emission filter in the wrong direction will adversely affect optical performance. Place the Olympus emission filter with the arrow ( ) on the side of the filter facing to the fluorescence cube. For other commercially available emission filters, refer to the instruction manuals.
  • Page 126 5. Secure the knob to the fluorescence cube. When installing to the NDD unit (reflected) (FV30-RXD, FV30-ARXD or FV30-AGAPD) Tighten the knob to the screw hole on the surface where the seal describing the product name is attached. When installing to the NDD unit (transmitted) (FV30-TXDBX) Tighten the knob to the screw hole at the bottom of the mount dovetail.

  • Page 127: Spectral Dm Cube (Fv30-Sdmopt)

    } Be careful not to soil the dichroic mirror surface dirty with sebum, etc. or scratch it with tools, etc. } Installing the dichroic mirror in the wrong direction will adversely affect optical performance. For Olympus dichroic mirrors, a corner of the interference film surface is marked with a marker. For other commercially available dichroic mirrors, refer to the instruction manuals of commercially available dichroic mirrors.

  • Page 128: Fluorescent Mirror Unit And Dm Unit (Maintenance)

    CAUTION falls under the category of service work. Olympus service personnel, or technical personnel who received service training, will carry out with their attachment to and removal from this system, in accordance with the FVMPE-RS Service Manual. Optical unit name...
  • Page 129 (toward the dichroic mirror frame), and fix it with the dichroic mirror fixing spring c . } If you are using an Olympus dichroic mirror, the reflection surface is the surface without a marker at the edge of the dichroic mirror.
  • Page 130 Place the excitation filter g on the fluorescence unit and fix it with the excitation filter fixing frame f . } If you are using an Olympus excitation filter, place the filter so that an arrow mark ( ) on the side faces the mirror unit.

  • Page 131: Dm Unit For The Sim Scanner (Fv30-Admopt)

    DM unit for SIM scanner, are supported by the Olympus service personnel. } If you use this instrument without registering the correction data, the problems, e.g. the image position is not matched, the resolution is deteriorated, etc.

  • Page 132: Visual Observation / Observation Using A Camera

    Visual observation / observation using a camera 6-1 Touch panel controller (Operation/Maintenance) Basic operations of touch panel controller (Operation/Maintenance) The touch panel controller can be operated by directly touching the displayed button with your fingertip. } The touch panel controller uses a static capacitance type touch panel and may not react if it is touched by a finger wearing a rub- ber or vinyl glove.

  • Page 133: Starting The Touch Panel Controller (Operation/Maintenance)

    FVMPE-RS Starting the touch panel controller (Operation/Maintenance) After settting the main switch of the touch panel controller to I (ON), the black startup display is displayed (for about 20 seconds), and then the blue startup display appears. } The order to turn ON the power switches of this system is defined. Refer to page 10 for details.

  • Page 134: System Setting (Maintenance)

    System setting (Maintenance) The system setup is required when using the system for the first time or after replacing a unit connected to the microscope. } The microscope cannot be controlled unless the unit setting and optical setting are completed. Be sure to set them. Tap the System Setting button.
  • Page 135 FVMPE-RS Optical setting This tab is used to set the optical system (objectives, mirror units and condenser). Objective setting Tap the [Optical] tab in the [System Setting] display to open the [Optical] display. Tap the [Objective Lens] tab in the [Optical] display. Register the objective actually mounted on the BX63LF for each nosepiece position.
  • Page 136 Condenser setting Tap the [Optical] tab in the [System Setting] display to open the [Optical] display. Tap the [Condenser] tab in the [Optical] display. Register the optical element actually mounted on the condenser for each condenser turret position number. Tap one of the turret posi- tion numbers in use to open the [Condenser] screen for the number.

  • Page 137: Selecting The Mode (Operation)

    FVMPE-RS Selecting the mode (Operation) Tap [Start Operation] in the [MENU] screen to display the following screen. } For detailed operating methods of the touch panel controller during observation, refer to Online Help for the touch panel controller. } To display the Online Help, tap Online Help button on the touch panel. Online Help button...

  • Page 138: Controller U-Mcz (Operation)

    6-2 Controller U-MCZ (Operation) Module nomenclature Observation mode selector buttons** FL SHUTTER button* Coarse adjustment knob Fine adjustment knob Detachable. Mirror unit switching button Switches over the fluorescence mirror units in the motorized cassette turret. Transmitted light brightness control buttons Adjusts the halogen bulb brightness.

  • Page 139: Fluorescence Observation (Reflected Light) (Operation)

    FVMPE-RS 6-3 Fluorescence observation (Reflected light) (Operation) Reflected fluorescence observation procedure } Operation parts names are displayed on the controller U-MCZ. } For operations by the touch panel controller, refer to Online Help of the touch panel controller. Preparation (Controls used) Main switch Set the main switch of the power supply unit to “...

  • Page 140: General Precautions For Observation

    } This care is not required when the U-FDICT transmitted DIC mirror unit with built-in analyzer is used. 3. Always use Olympus immersion oil for oil immersion objectives. 4. If you use an objective with correction collar, you can correct contrast degradation due to variation in cover glass thick- ness by adjusting the correction collar.

  • Page 141: Selecting The Fluorescent Mirror Unit Cassette

    FVMPE-RS Selecting the fluorescent mirror unit cassette Selecting the position • U-MCZ See page 66. • Touch panel controller. Refer to Online Help for the touch panel controller. • FV30S-SW Refer to Online Help for FV30S-SW. Observation method Position LSM observation Reflected light fluorescence observation by mercury lamp Fluorescent mirror unit for each excitation wavelength...

  • Page 142: Preparing Transmitted Observation (Maintenance) (Bx63Lf Only)

    Preparing transmitted observation (Maintenance) (BX63LF only) Attaching the polarizer (Maintenance) } If FV30-TXDBX is combined with the system, the filter turret cannot be combined. Drop in the filter frame reinforcing ring provided with BX63LF into the filter holder on the microscope frame. Loosen filter turret clamping screws using the provided Allen wrench.

  • Page 143: Transmitted Brightfield Observation (Operation) (Bx63Lf Only)

    FVMPE-RS Transmitted brightfield observation (Operation) (BX63LF only) Flow of brightfield observation } Operation parts names are displayed on the controller U-MCZ. } For operations by the touch panel controller, refer to Online Help of the touch panel controller. Preparation (Controls used) Change the observation mode to the bright field Observation mode selector buttons •...

  • Page 144: Differential Interference Contrast (Dic) Observation (Operation/Maintenance)

    CAUTION category of service works. Olympus service personnel or engineers who received the service training carry out the services in accordance with “FVMPE-RS Service Manual”. } The normal optical performance of DIC observation cannot be achieved if a plastic petri dish is used.
  • Page 145 FVMPE-RS Remove the condenser from the microscope. Remove an objective and engage the hole without the objective in the light path. Engage the polarizer and analyzer in the light path and turn the transmitted light on. Remove the eyepiece from the eyepiece sleeve, look into the sleeve, turn the polarizer rotation dial so that the black interference stripe is darkest, and tighten the clamping polarizer knob } The interference stripe is not visible clearly if the field iris is focused insufficiently...
  • Page 146 } With WI-UCD or WI-DICD, follow the procedures described below. Turn the quarter-wave plate rotation ring so that the black interference stripe seen at the center of the eyepiece sleeve’s field of view, then tighten the clamping knob . Ignore the short interference stripes in the surroundings in this adjustment.

  • Page 147: Observation Method (Operation)

    FVMPE-RS Observation method (Operation) Engage the objective to be used in the light path. Engage the DIC prism of the nosepiece in the light path. If you are using BX63L-DSGNPS or BX63L-DSLNPS, push in the DIC prism IN/OUT knob If you are using BX63L-D6RES, push in the slider. } If you are using BX63-DSLNPS, move the DIC prism IN/OUT knob while holding the objective selector knob to prevent the objective...

  • Page 148: Lsm Observation (Operation)

    LSM observation (Operation) } For details, refer to the Online Help for FV30S-SW. } When using this system, we recommend turning the indoor lighting low in order to reduce external distur- bance light on observations. (The indoor lighting particularly affects the differential interference observation images.) } Do not apply cool air to this system from the air conditioner or so.

  • Page 149: Signal Connection With External Device

    FVMPE-RS Signal connection with external device With the external device detecting unit FV30-ANALOG, this system is able to transmit and receive electric signals to and from external devices. 8-1 Features of FV30-ANALOG • Capable of receiving electric signals transmitted from external devices as an image equivalent to that acquired via four channels •...
  • Page 150 Items Specifications Analog voltage Sampling rate This information will be a data for Sampling specifications acquiring the latest sampling value at Galvano scan mode: 1 Msps each pixel timing in each scan mode. Resonant scan mode: 3.3 Msps Dip switch for changing the analog voltage input range The input range settings by the dip switch are...
  • Page 151 FVMPE-RS Items Specifications Signal ground To be connected to the chassis ground. Connection of this output to external mechanism In order to prevent the system from malfunctions such as noises, etc., use the Schmitt trigger type input IC for the logic IC, etc. to which this signal is inputted. Input impedance of the external mechanism connected to this output 10 kΩ...
  • Page 152 Items Specifications Trigger output Output channel number: 6 ch specifications Since OUT6 is a reserved port, it cannot be used with this system. Output signal level TTL signal level Output voltage L: 0.4 V or lower Output voltage H: 2.7 V or higher Signal ground To be connected to the chassis ground.

  • Page 153: Setting Procedure

    FVMPE-RS 8-3 Setting procedure Analog input connector Trigger input connector Synchronization signal output connector Trigger output connector TRAC1(MAIN) TRG IN SCAN LOOP PCLK TRAC2(SIM) TRG OUT OUT1 OUT2 OUT3 OUT4 OUT5 OUT6 SCAN LOOP PCLK ANALOG IN ANALOG OUT TRAC3 TRAC4 O A B C ON O A B C...

  • Page 154: Connection

    Connection 1. Set the analog input range after turning OFF the powers of FV30-ANALOG and external devices. } The power of FV30-ANALOG is automatically turned OFF when the system power (FV30-PSU) is turned OFF. 2. The cables used for connection must meet following specifications. •...

  • Page 155: Specifications Of Synchronizing Signal Output Timing

    FVMPE-RS 8-4 Specifications of synchronizing signal output timing Synchronizing Signal Type There are 6 types of synchronizing signals outputted from FV30-ANALOG. SCAN This signal indicates that the scan is being performed. This is a TTL signal where Scanning = H level and Being stopped = L level.

  • Page 156: Synchronizing Signal Output Timing By Scan Mode

    Synchronizing Signal Output Timing by Scan Mode The relation between the scan operation and the synchronizing signal output in each mode is illustrated below. XY(Region) scan Scanner trajectory Imaging (or stimulation) zone Start SCAN 1 Line 1 Frame Last frame Resonant XY scan Scanner trajectory Imaging zone...
  • Page 157 FVMPE-RS Free line scan Scanner trajectory Imaging (or stimulation) zone Start SCAN 1 Line Point scan Scanner trajectory Imaging (or stimulation) zone Start SCAN Point Multi point scan Scanner trajectory Imaging (or stimulation) zone Start SCAN 1 Cycle Tornado scan Stimulation zone Start SCAN...

  • Page 158: Loop Signal Output Timing

    LOOP Signal Output Timing The LOOP signal output timing in each series scan is illustrated below with operation examples. Live scan Note that the waveshape of Live scan in Line-Z is same as that in Line ZT illustrated below Start Live Stop Live Stop...
  • Page 159 FVMPE-RS XYZ scan Start Start SCAN LOOP XYZT scan Z=7, T=2 Start Start SCAN LOOP Line ZT scan Start Start SCAN 1HD Width LOOP...

  • Page 160: Preventive Inspection Sheet For Illumination Devices

    · If there are any check marks ( ) noted, immediately stop use of the product and seek service or replacement for the illumination device(s) for prevention. · If you detect an abnormality other than that listed below with your illumination device or other Olympus product, request inspection from your Olympus distributor.

  • Page 161: Care And Storage (Maintenance)

    · In high-humidity environment, condensation may occur in this system. So follow the environmental condition not only in use but also in storage for a long term. In addition, there is a desiccant agent in the FV30-AGAPD to prevent the appearance of condensation. If you want the replacement, please contact your local Olympus sales office. (charged)

  • Page 162: Troubleshooting Guide (Operation/Maintenance)

    Depending on usage, this system may not deliver the expected performance despite not being at fault. Should any problem occur, refer to the following list to take the appropriate corrective action. If a problem persists, contact your local Olympus sales office. 11-1 LSM observation...
  • Page 163 4. The image is disturbed. There is too much vibration at the location Consult with your local Olympus sales office. where this system is installed. External light such as that from a fluores- Darken the room before acquiring the cent lamp is detected.
  • Page 164 Replacing the GaAsP-PMT with a new one dark and contains much (paid) is needed, please contact your local noise. Olympus sales office. It is a consumable product. The laser light is weak. Increase the laser intensity. The selected excitation wavelength is not...

  • Page 165: Visual Observation

    FVMPE-RS 11-2 Visual observation Problem Cause Remedy a) The halogen bulb does not light. Power cord of the BX63L-CBH is Plug the power cord into a power unplugged. outlet. Main switch of the BX63L-CBH is not Set the main switch to “ I ” (ON). “...
  • Page 166 Front lens of the objective is dirty. Clean the objective. The immersion oil appropriate with an Use Olympus immersion oil with the oil immersion objective is not used. oil immersion objective, Olympus silicone oil with the silicone immersion objective and water with the water immersion objective.
  • Page 168 Manufactured by Shinjuku Monolith, 2-3-1 Nishi-Shinjuku, Shinjuku-ku, Tokyo 163-0914, Japan Distributed by 48 Woerd Avenue Waltham, MA 02453, U.S.A. 8F Olympus Tower, 446 Bongeunsa-ro, Gangnam-gu, Seoul, 06153 Korea AX8334 09...

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