Nikon TIRF2 Instructions Manual
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Nikon TIRF2 Instructions Manual

Nikon TIRF2 Instructions Manual

System for te2000
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M339E
04.12.NF.2
TIRF2 SYSTEM FOR TE2000
INSTRUCTIONS

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  • Page 1 M339E 04.12.NF.2 TIRF2 SYSTEM FOR TE2000 INSTRUCTIONS...
  • Page 3 Before using the system, carefully read the precautions concerning correct use provided at the beginning of this manual. • No part of this manual may be reproduced or transmitted in any form without the prior consent of Nikon. • The contents of this manual are subject to change without notice.
  • Page 4 WARNING and CAUTION Symbols Although Nikon products are designed to provide the utmost safety during use, incorrect usage or failure to follow the safety instructions provided may cause personal injury or property damage. Damage or injury that may occur due to mishandling is unwarranted.
  • Page 5 IEC Class 2 Laser Product 2. Intended product use This product may be used only for microscopic observations. It must be mounted to the specified Nikon microscope model TE2000-E/U/S. It may not be used for any other purpose. 3. Setting up this product Setup of this product must be handled by trained service personnel.
  • Page 6 Using other than the specified type of lamp could result in an accident, such as a burst. Because safety is a top priority in the design of Nikon products, the hazards described above should not pose any danger as long as you heed all of the warnings and cautions in the manuals and use the system only for its intended purpose.
  • Page 7 If water enters a component, immediately suspend use of this product, disconnect the power cord from the outlet, and contact your nearest Nikon representative. 6. Do not place any objects on system equipment To prevent deformation, failure, or malfunction, and to avoid injury, never place any objects (heavy ones in particular) on any component of the system.
  • Page 8 • Make sure that this product is not subject to strong vibrations, which may degrade image quality. Consult with your Nikon representative about preventive measures you may take when this product is installed.

  • Page 9: Table Of Contents

    29 TIRF microscopy ....................30 Termination tasks....................32 Part Descriptions..................33 TIRF2 attachment ....................33 Specimen cover ..................... 38 Eyepiece shutter ....................39 Shutter control unit (T-LUSU, T-LUSU2) ..............40 T-FLC-HQ cassette holder ..................42 Laser unit......................
  • Page 10 Contents Disinfecting the product ..................56 Storage......................... 56 Moving and relocating the system (charged).............. 56 Periodical inspection (charged) ................56 VIII Performance and Specifications ..............57 Overall configuration ....................57 Shutter control unit ..................60 AC adapter for shutter control unit ..............60 Power cord for AC adapter of shutter control unit ..........

  • Page 11: System Configurations And Names

    (8) Eyepiece shutter (9) Cassette holder (10) Power supply for microscope (11) TIRF2 attachment (12) Light source for the Epi-illumination attachment (Super high pressure mercury lamp housing is indicated in the figure.) (13) Power supply of the light source for the Epi-illumination attachment (Power...
  • Page 12 (8) Eyepiece shutter (9) Cassette holder (10) Power supply for microscope (11) TIRF2 attachment (12) Light source for the Epi-illumination attachment (Super high pressure mercury lamp housing is indicated in the figure.) (13) Power supply of the light source for the Epi-illumination attachment (Power...

  • Page 13: Laser Path

    System configurations and names 2 Laser path Laser path Laser unit Two-laser unit Three-laser unit Three-laser unit C-LU2 C-LU3 C1-LU3 (1) Laser mounting position L1 (See table below.) (2) Laser mounting position L2 (See table below.) (3) Laser mounting position L3 (See table below.) (4) Motorized shutter (5) ND filter (ND4, ND8) (6) Laser synthesis mirror...

  • Page 14: Microscope Part

    System configurations and names 2 Laser path Microscope part TE2000 MICROSCOPE Laser beam TE2000 MICROSCOPE (from top) Laser beam...

  • Page 15: Safety Labels

    System configurations and names 3 Safety labels Safety labels (1) IEC laser warning label - (2) IEC explanatory label (3) Model name and serial hazard symbol Class 2 laser product number label (4) Safety standard label (5) IEC caution label- (6) IEC caution label- Class 3B areas Class 2 areas...
  • Page 16 System configurations and names 3 Safety labels TE2000 (1) IEC laser warning label - (2) IEC explanatory label hazard symbol Class 2 laser product (Labels on stage)

  • Page 17: Two-Laser Unit C-Lu2

    System configurations and names 3 Safety labels Two-laser unit C-LU2 (1) IEC laser warning label - (5) IEC caution label - hazard symbol Class 3B areas...

  • Page 18: Three-Laser Unit C-Lu3

    System configurations and names 3 Safety labels Three-laser unit C-LU3 (1) IEC laser warning label - (5) IEC caution label - hazard symbol Class 3B areas...

  • Page 19: C1-Lu3

    System configurations and names 3 Safety labels Three-laser unit C1-LU3 (1) IEC laser warning label - (5) IEC caution label - hazard symbol Class 3B areas...

  • Page 20: Shutter Control Unit (T-Lusu, T-Lusu2)

    System configurations and names 3 Safety labels Shutter control unit (T-LUSU, T-LUSU2) (4) Safety standard label (Back) This label is used on both of the T-LUSU and T-LUSU2.

  • Page 21: Tirf2 Attachment

    System configurations and names 3 Safety labels TIRF2 attachment (3) Model name and serial (6) IEC caution label- (7) IEC caution label- number label Class 2 areas Class 2 areas...

  • Page 22: Names Of Component Parts

    System configurations and names 4 Names of component parts Names of component parts Microscope part Eyepiece shutter Bino : Eyepiece shutter open (for Epi-fl or bright-field microscopy) Confocal : Eyepiece shutter close (for TIRF microscopy) Set the dial to "Confocal" for TIRF observation.
  • Page 23 System configurations and names 4 Names of component parts Specimen cover TIRF2 attachment (10) (12) (14) (13) (16) (15) (11) (16) (11) (12) (10) (15) (13) (14) Parts for TIRF microscopy operation Parts for Epi-fl microscopy operation (1) Optical fiber...

  • Page 24: Laser Unit

    System configurations and names 4 Names of component parts Laser unit Two-laser unit (C-LU2) Laser mounting position L1 Laser mounting position L2 Shutter control unit connector ND filter slider Single-mode fiber C-LU2 Three-laser unit (C-LU3, C1-LU3) Laser mounting position L1 Laser mounting position L2 Laser mounting position L3 Shutter control unit connector...

  • Page 25: Rough Sequence Of Microscopy

    Rough Sequence of Microscopy Preparation before Center the Epi-illumination lamp. (It should be executed each time microscopy you replace the Epi-illumination lamp.) Adjust the diopter and the interpupillary distance. Safety check A TV camera or dust-proof cap is in place on each of the ports on the microscope.

  • Page 26: Detailed Sequence Of Microscopy

    For the Epi-fl attachment using the mercury lamp as the light source for Epi-fl microscopy, centering is required each time the lamp is replaced. See the manual “Nikon Super High Pressure Mercury Lamp Power Supply” or “High-intersity Light Source” to center the lamp.

  • Page 27: Startup Tasks

    Detailed Sequence of Microscopy 2 Startup tasks Startup tasks Emitting the laser (1) Emitting the air-cooled argon laser (1) Switch on the power switch. 1 Switch on the power switch and turn the key switch clockwise on the air-cooled argon laser power unit to turn on the unit.

  • Page 28: Turning On Power To Other Components

    Detailed Sequence of Microscopy 2 Startup tasks (3) Setting-in the ND filter To prevent the intensity of the laser from degrading the sample, we recommend inserting an ND filter (ND4 or ND8) into the optical path in advance. ND filter Turning on power to other components Turn on the microscope, Epi-illumination lamp, shutter control unit, camera etc.

  • Page 29: Epi-Fl Microscopy (Bring The Specimen Into Focus)

    All the laser shutters are automatically closed. (2) Change the optical path. Select the Epi-fl microscopy. (Push in the mirror switch lever on the TIRF2 attachment.) Set the optical path of the Set the dial to "Bino" for normal microscope to “observation 100%”.
  • Page 30 Detailed Sequence of Microscopy 3 Epi-fl microscopy (bring the specimen into focus) (6) Center the field diaphragm and adjust its size. Field diaphragm Reduce the field diaphragm opening. (Move the field open/close lever diaphragm open/close lever to the “C” (close) direction.) Move the center of the field diaphragm image to the center of the viewfield.

  • Page 31: Simple Surface Reflection Interference Microscopy Using Epi-Fl Attachment (Checking The State Where The Specimen Contacts To The Cover Glass)

    Detailed Sequence of Microscopy 4 Simple surface reflection interference microscopy using Epi-fl attachment (checking the state where the specimen contacts to the cover glass) Simple surface reflection interference microscopy using Epi-fl attachment (checking the state where the specimen contacts to the cover glass) In theory, it is possible to make high-contrast observations of areas in contact with the cover glass using TIRF illumination.

  • Page 32: Tirf Microscopy

    Set the dial to "Bino" for normal observation. (Eyepiece shutter open) (3) Change the optical path. Select the TIRF microscopy. (Pull out the mirror switch lever on the TIRF2 attachment.) Switch the microscope’s optical path to a port to which the TV camera is attached.
  • Page 33 Detailed Sequence of Microscopy 5 TIRF microscopy (6) Collimate the laser beam (Perform each time the objective is changed). WARNING Never attempt to look into the laser beam. (FDA CDRH: Class II, EN60825-1: Class 2, IEC60825-1: Class 2) The objective emits a powerful laser beam. When adjusting the laser position adjustment knob, never attempt to look into the objective or the areas near the objective from above or below the stage as the lens emits a powerful laser beam toward the top and bottom sides of the stage.

  • Page 34: Termination Tasks

    Detailed Sequence of Microscopy 6 Termination tasks Termination tasks First of all, turn off the laser. 1） Turn off the laser. See the Instruction Manual supplied with the laser before turning off the laser, and unplug the power cord. • For air-cooled argon laser Switch off the key switch on the air-cooled argon laser remote controller, and leave it for approximately 15 minutes to sufficiently cool down the laser head.

  • Page 35: Part Descriptions

    Part Descriptions TIRF2 attachment TIRF2 attachment is the main optical unit of this system. Excitation light from a laser unit is guided into this unit through an optical-fiber cable. Excitation light emitted by this unit forms an image on the pupil surface of the objective.
  • Page 36 Part Descriptions 1 TIRF2 attachment (1) Optical-fiber cable This cable conducts laser beam from the laser unit to the TIRF2 attachment. CAUTION • To ensure specified performance and to protect your eyes and skin from exposure to the laser, do not detach the optical-fiber cable from the TIRF2 attachment.
  • Page 37 Part Descriptions 1 TIRF2 attachment (6) Indices (7) Condenser lens locking screw (8) Focal point adjustment knob You can change the focal point of the laser beam along the path of the objective's optical axis by moving this knob back and forth.
  • Page 38 Part Descriptions 1 TIRF2 attachment (9), (10) Field diaphragm (Field diaphragm centering adjustment screws, Field diaphragm open/close lever) The field diaphragm is adjusted by using the field diaphragm centering adjustment screws and the field diaphragm open/close lever. The field diaphragm restricts illumination to the area of the specimen that is being viewed.
  • Page 39 Part Descriptions 1 TIRF2 attachment (11), (12) Aperture diaphragm (Aperture diaphragm centering adjustment screws, Aperture diaphragm open/close lever) The aperture diaphragm is adjusted by using the aperture diaphragm centering adjustment screws and the aperture diaphragm open/close lever. The aperture diaphragm is used to adjust the numerical aperture of an illumination optical system.

  • Page 40: Specimen Cover

    Part Descriptions 2 Specimen cover Specimen cover This cover employs a filter that can attenuate the intensity of light in the proximity of 488 nm and 532 nm wavelengths by a factor of up to 1000. Make sure that the cover is always in place, except when you are adjusting the laser focal point.

  • Page 41: Eyepiece Shutter

    (observation using eyepieces) so that the laser beam is not guided into the TIRF2 attachment. An illumination pillar switch to be mounted on the microscope's illumination pillar is supplied with the eyepiece shutter.

  • Page 42: Shutter Control Unit (T-Lusu, T-Lusu2)

    Part Descriptions 4 Shutter control unit (T-LUSU, T-LUSU2) Shutter control unit (T-LUSU, T-LUSU2) This unit is designed to open or close the motorized shutter housed within each laser unit. Although the unit can open or close up to three channels of motorized shutters, operation of motorized shutters is conducted through mutually exclusive control, as a result of which only one shutter can be opened at a time.
  • Page 43 Part Descriptions 4 Shutter control unit (T-LUSU, T-LUSU2) (1) Power switch Turns the 12 VDC power from the AC adapter on or off. (2) Power lamp Lights green when the power is on. (3) AC adapter connector Connects the AC adapter output. (4) Eyepiece shutter connector Connects the microscope-mounted eyepiece shutter connector.

  • Page 44: Flc-Hq Cassette Holder

    Filter cubes that incorporate dichroic mirrors are attached in the T-FLC-HQ cassette holder. The filter cube reflects the laser beam or Epi-fl illumination from the TIRF2 attachment to guide it to the objective. Up to six filter cubes can be attached and replaced as necessary.
  • Page 45 Part Descriptions 5 T-FLC-HQ cassette holder Replacing filter cubes: "Address on the optical path" Detach the filter cube port cover, check the marking (area turret address, align the filter cube with the Excitation method where a sticker guide groove, and insert the cube. switching ring is to be affixed) Turn the excitation method changeover ring...
  • Page 46 Part Descriptions 5 T-FLC-HQ cassette holder When self-fluorescence is pronounced, use an excitation filter with a narrow bandwidth. (The resulting fluorescent image will be darker, however.) Excitation filters are likely to deteriorate the more they are used, since they are exposed to intense light. Replace excitation filters as necessary.
  • Page 47 Part Descriptions 5 T-FLC-HQ cassette holder BP filters (band-pass filters) A BP filter allows only light in a certain range of FITC fluorescent wavelengths to pass. wavelength band This type of filter is used to view the fluorescent image BA520-560 created by a specific stain when a given specimen has (BP type) multiple stains.

  • Page 48: Laser Unit

    6 Laser unit Laser unit The laser unit is designed to introduce the laser beam into the TIRF2 attachment through a single-mode fiber. The unit is equipped with three laser units: one two-laser unit and two three-laser units. Connect the fiber to one of the three.
  • Page 49 Part Descriptions 6 Laser unit Mountable position Two laser unit Three laser unit Three laser unit Product Remarks C-LU2 C-LU3 C1-LU3 An Ar wavelength selector Spectra-physics Model 163-C02 is required for use. (488-514) An Ar wavelength selector Spectra-physics Model 161C-02 is required for use.

  • Page 50: Shutter Control Unit Communications Commands

    Shutter Control Unit Communications Commands Communication-controlled operation You can control this system from your PC or other devices through the RS-232C interface. Create your own control program based on the information given in this manual. 1-1 Communications system Start-stop serial communication 1-2 Connector specification The following signals are used with the system's serial interface connector: D-sub 9-pin connector (male)
  • Page 51 Shutter Control Unit Communications Commands Communication-controlled operation 1-4 Communications parameters Baud rate: 9600 bps (fixed) Data length: 8 bits (fixed) Start bits: 1 bit (fixed) Stop bits: 1 bit (fixed) Parity check: None 1-5 Communications format The data format used when the system receives data from external equipment is referred to as “command format,”...
  • Page 52 (five commands) Processing inhibit Processing is inhibited error Processing stop Command processing is stopped error 1-6 Communications sequence [TIRF2 system] ［PC］ c command Command processing o or n response [TIRF2 system] ［PC］ r command Command processing a or n response...
  • Page 53 Shutter Control Unit Communications Commands Communication-controlled operation 1-7 Commands c command: Operation, control, and data setting command o: Normal response c command Description of operation n: Error response Opens a specified shutter and closes any other shutters. If an open time period has been specified, the shutter will close when the specified period elapses.

  • Page 54: Troubleshooting Tables

    Replace the lamp. of its operational life. If the fuse can be replaced, replace The fuse is blown. it. Otherwise, contact your nearest Nikon representative. The shutter is in the optical Remove the shutter from the optical path. path. Rotate the excitation method...
  • Page 55 NCG objective.) No immersion oil has been applied to the tip of an Apply Nikon Immersion Oil DF. Viewing is poor. immersion-oil type objective. The specified immersion oil is Apply Nikon Immersion Oil DF.
  • Page 56 Troubleshooting Tables TIRF microscopy Problem Cause Remedy Unable to turn on system The power cable is not Connect the power cable to the power connected to the outlet. outlet. Turn on the laser in accordance with the steps described in "2. startup The laser is not on.

  • Page 57: Care And Maintenance

    Care and maintenance WARNING Do not remove any components or parts on the system. Never attempt to remove the objective, filter cube, optical fiber, lamphouse, eyepiece lens, and any other parts during operation. Removing any one of them may cause the laser beam to be emitted through that portion, resulting in exposure of eyes or skin to the laser.

  • Page 58: Cleaning The Product

    • Detach the laser from the laser unit. • Detach the optical-fiber cable from the laser unit. • Detach the optical-fiber cable from the TIRF2 attachment. • Detach the TIRF2 attachment from the microscope. • Detach the cassette holder from the microscope.

  • Page 59: Performance And Specifications

    When connecting a optical fiber that has a straight tip, install a straight fiber adapter to the TIRF2 attachment. Stageup lens When using the TIRF2 attachment on the lowest step during stageup, install the stageup lens to the TIRF2 attachment. T-BSUK70 70mm upkit and T-FLS FL spacer are required to stageup.
  • Page 60 VIII Performance and Specifications 1 Overall configuration Table 1 Mountable Lasers Color Product Specification Mountable unit (position) Multi Lines Manufacturer: Spectra-physics 488-514nm/40mW C-LU2(L1) Head: 163-C02 C-LU3(L2) Power supply: 263-C04 C1-LU3(L1) Manufacturer: Spectra-physics 488-514nm/25mW C-LU2(L1) Head: 161C-02 C-LU3(L2) Power supply: 263-C04 C1-LU3(L1) Violet Manufacturer: Coherent...
  • Page 61 VIII Performance and Specifications 1 Overall configuration Color Product Specification Mountable unit (position) Manufacturer: JDS Uniphase 633nm/15mW C-LU2(L2) Head: 1144P C-LU3(L1) Power supply: 1218-3/100V, C1-LU3(L3) 1218-1/115V, 1218-2/230V Manufacturer: JDS Uniphase 633nm/10mW C-LU2(L2) Head: 1135P C-LU3(L1) Power supply: 1216-3/100V, C1-LU3(L3) 1216-1/115V, 1216-2/230V Manufacturer: JDS Uniphase 633nm/5mW C-LU2(L2)

  • Page 62: Shutter Control Unit

    VIII Performance and Specifications 1 Overall configuration Shutter control unit Type T-LUSU or T-LUSU2 Input rating 12 VDC 1A AC adapter for shutter control unit Manufacturer PHIHONG ENTERPRISE (Taiwan) Type PSA-30U-120（N） Input rating 100-240 VAC 0.7A 50/60Hz Voltage fluctuation ±10% Output rating 12 VDC Rated output current...

  • Page 63: Compatible Standards

    VIII Performance and Specifications 1 Overall configuration Compatible standards Laser safety • This product complies with IEC60825-1 Class 2 requirements. • This product complies with FDA Class II requirements. CE marking • This product meets EU Low Voltage Directive requirements. •...

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