Zeiss TIRF 3 Operating Manual
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Zeiss TIRF 3 Operating Manual

Zeiss TIRF 3 Operating Manual

Laser imaging system
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Summary of Contents for Zeiss TIRF 3

  • Page 2 Carl Zeiss Copyright Laser TIRF 3 Knowledge of this manual is required for the operation of the instrument. You should therefore familiarize yourself with the contents of this manual and pay special attention to instructions concerning the safe operation of the instrument.

  • Page 3: Table Of Contents

    Laser TIRF 3 Contents Carl Zeiss CONTENTS Page DEVICE SAFETY ...................... 6 General Notes ........................6 Safety Instructions ......................8 1.2.1 Laser Radiation ........................8 1.2.2 Installation Instructions ....................... 9 1.2.3 Notes on Computer and Software ..................11 1.2.4 Service and Customer Interfaces ..................12 1.2.5...
  • Page 4 Carl Zeiss Contents Laser TIRF 3 Mounting the Laser Safety Cover for Reflected Light Provided for Mechanical and Scanning Stages ........................57 Mounting the S TIRF S1/S TIRF DARK S1 Incubator .............58 Incubator XLmulti S1 RED LS and Incubator XLmulti S1 DARK LS.........60 3.6.1...
  • Page 5 Laser TIRF 3 Contents Carl Zeiss Mounting the Camera ...................... 89 Connecting the HXP 120 Illumination Unit ................ 90 Service Request ........................ 91 USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS ....92 Laser Safety Concept ......................93 Safety Kit for Customer-Provided Laser Module ..............95 Remote switch for laser mode ...................

  • Page 6: Device Safety

    DEVICE SAFETY Carl Zeiss General Notes Laser TIRF 3 DEVICE SAFETY General Notes The Laser TIRF imaging system has been designed, produced and tested in compliance with the standards DIN EN 61010-1 (IEC 61010-1) "Safety requirements for electrical equipment for measurement, control and laboratory use", DIN EN 60825-1 (IEC 60825-1) "Safety of laser equipment", and DIN EN 61326-1...
  • Page 7 DEVICE SAFETY Laser TIRF 3 General Notes Carl Zeiss In addition to this manual of the Laser TIRF imaging system, please observe the detailed information given in the following operating manuals: − Operating manual for the Axio Observer microscope (B 46-0111) −...

  • Page 8: Safety Instructions

    DEVICE SAFETY Carl Zeiss Safety Instructions Laser TIRF 3 Safety Instructions 1.2.1 Laser Radiation The Laser TIRF system is a class 3B laser system and labeled accordingly (total output power on the microscope: max. 500 mW). If you use laser instruments, adhere to the provisions concerning health protection and industrial safety.

  • Page 9: Installation Instructions

    Installation Instructions The Laser TIRF imaging system must be installed and commissioned by technicians trained in laser safety and authorized by Carl Zeiss. The client is liable to pay the costs of services rendered by the Carl Zeiss Microscopy GmbH.
  • Page 10 Carl Zeiss Microscopy GmbH or by persons expressly authorized by this company. This applies in particular to the microscope system, the Slider Laser TIRF, the Laser Bench Tower or Laser Bench Rack, the control PC system and the cable connections.

  • Page 11: Notes On Computer And Software

    To ensure proper functioning of the Laser TIRF imaging system, it should be subjected to an annual safety inspection. The safety checks must be carried out by an authorized Carl Zeiss service technician. All national safety checks should be carried out.

  • Page 12: Service And Customer Interfaces

    Service interfaces between Laser Bench Tower and Slider Laser TIRF CAUTION The fiber mounts (Fig. 1-1/1 and 2) may be detached by Carl Zeiss Service staff only. If one fiber end is not firmly connected to the fiber mount, laser class 3B radiation will emerge!
  • Page 13 (Fig. 1-2/2) and on the optional Laser TIRF (Fig. 1-2/3) and Cell Observer SD (Fig. 1-2/4) slider units may only be loosened by Carl Zeiss service technicians. If one fiber end is not firmly connected to the fiber mount, laser class 3B radiation will emerge!
  • Page 14 CAUTION The screws (Fig. 1-3/1) on the retaining ring (Fig. 1-3/2) for the camera adapter should only be unscrewed by Carl Zeiss service technicians! Non-observance involves the risk of eye damage! If no camera is mounted on the sideport, it should be sealed by the protective cover (Fig.
  • Page 15 DEVICE SAFETY Laser TIRF 3 Safety Instructions Carl Zeiss Fig. 1-4 Camera adapter - camera / protective cover customer interface The camera adapter (Fig. 1-4/1) is equipped with an interlock connection (two reflected light barriers) which automatically closes the laser shutter as soon as the camera (Fig. 1-4/2) or the protective cover (Fig.
  • Page 16 CAUTION The screws (Fig. 1-5/1) on the sensor ring (Fig. 1-5/2) and the screw (Fig. 1-5/6) on the contact ring (Fig. 1-5/4) may only be unscrewed by Carl Zeiss service technicians! Non-observance involves the risk of eye damage! Service interfaces for HXP 120 illumination unit...
  • Page 17 Transmitted light carrier - HAL 100 Illuminator service interface CAUTION The clamp screw (Fig. 1-6/1) is a safety screw and may only be unscrewed by Carl Zeiss Service technicians. If the lamp housing is removed, laser class 3B radiation may emerge!

  • Page 18: Warning And Information Labels On The Instrument

    Observe the warning and information labels affixed to the Laser TIRF imaging system and Axio Observer microscope. Please check that all labels shown below are actually in place. Contact Carl Zeiss Microscopy GmbH if a sign or label is found to be missing. You will receive replacements free of charge.
  • Page 19 DEVICE SAFETY Laser TIRF 3 Safety Instructions Carl Zeiss Fig. 1-8 Warning and information labels on the Laser TIRF imaging system with laser safety cover (for labels on the microscope stand see Fig. 1-7) Fig. 1-9 Warning and information labels on the Slider Laser TIRF...
  • Page 20 DEVICE SAFETY Carl Zeiss Safety Instructions Laser TIRF 3 Fig. 1-10 Warning and information labels on the Laser Bench Tower 423683-7144-001 02/2014...
  • Page 21 DEVICE SAFETY Laser TIRF 3 Safety Instructions Carl Zeiss Fig. 1-11 Warning and information labels on the Laser Bench Rack / 3F Fig. 1 Warning and information labels on the SVB 1 02/2014 423683-7144-001...

  • Page 22: Notes On Warranty

    Repairs are only permitted to be carried out by Carl Zeiss Service staff or persons especially authorized by it. If any defects occur with the instrument, first contact your local Carl Zeiss representative.

  • Page 23: Instrument Description

    INSTRUMENT DESCRIPTION Laser TIRF 3 Intended Use Carl Zeiss INSTRUMENT DESCRIPTION Intended Use Product name: Laser TIRF imaging system The Laser TIRF imaging system is used to visualize fluorescence-capable substances / molecules (excitable by light) by means of an electromagnetic (evanescent) field generated by total reflection. Typically, this field is generated at the transition from the cover slip to the medium and decays exponentially with the distance (see Section 2.2 on page 24).

  • Page 24: Operating Principle

    INSTRUMENT DESCRIPTION Carl Zeiss Operating Principle Laser TIRF 3 Operating Principle The analysis of images obtained with conventional wide-field fluorescence excitation is often difficult because of background fluorescence emitted in out-of-focus planes. This is due to the comparatively low Z resolution achievable with this illumination technique.
  • Page 25 INSTRUMENT DESCRIPTION Laser TIRF 3 Operating Principle Carl Zeiss If the angle Φ exceeds a critical value, total reflection will occur (Fig. 2-1/B). As Φ is 90° at this point, the critical angle Φ φ If the light, for instance, makes the transition from the cover slip (n = 1.518) to water (n = 1.33), the resulting critical angle will be 61°.

  • Page 26: Instrument Description And Main Features

    INSTRUMENT DESCRIPTION Carl Zeiss Instrument Description and Main Features Laser TIRF 3 Instrument Description and Main Features The following components are necessary for operating the Laser TIRF imaging system in combination with Axio Observer: − Microscope Axio Observer.Z1 (431007-9901-000 and higher) with basic laser safety kit, version 2, −...
  • Page 27 INSTRUMENT DESCRIPTION Laser TIRF 3 Instrument Description and Main Features Carl Zeiss Focus rotary button Active laser status LED TIRF divergence scale Light emergence aperture Safe laser status LED TIRF angle adjusting screw Fig. 2-2 Structure of Slider Laser TIRF man. / mot.

  • Page 28: Laser Bench Tower And Laser Bench Rack Or Laser Bench Rack 3F

    INSTRUMENT DESCRIPTION Carl Zeiss Instrument Description and Main Features Laser TIRF 3 2.3.2 Laser Bench Tower and Laser Bench Rack or Laser Bench Rack 3F The laser units for the individual laser lines and the corresponding electronics are housed either in the Laser Bench Tower or in the Laser Bench Rack depending on the equipment version selected.

  • Page 29: Signal Distribution Box

    INSTRUMENT DESCRIPTION Laser TIRF 3 Instrument Description and Main Features Carl Zeiss 2.3.3 Signal Distribution Box The system components are connected to the signal distribution box (see operating manual SVB 1). Fig. 2-5 Signal distribution box 2.3.4 TIRF Controller The motorized Slider Laser TIRF is controlled via the TIRF controller. The LEDs at the front indicate the operational status of the Slider Laser TIRF.

  • Page 30: Laser Safety For The Sample Compartment

    INSTRUMENT DESCRIPTION Carl Zeiss Laser Safety for the Sample Compartment Laser TIRF 3 Laser Safety for the Sample Compartment The laser light can escape from the objective at different angles, meaning that laser safety must be ensured for the sample compartment.

  • Page 31: Basic Kit Laser Safety Rev. 2

    INSTRUMENT DESCRIPTION Laser TIRF 3 Basic kit laser safety Rev. 2 Carl Zeiss Basic kit laser safety Rev. 2 The basic kit laser safety Rev. 2 includes an eyepiece shutter (factory-installed or retrofitted by service personnel), a laser safety board for the stand, a distribution box and sticker labels.

  • Page 32: Remote Switch For Laser Mode

    INSTRUMENT DESCRIPTION Carl Zeiss Remote switch for laser mode Laser TIRF 3 Remote switch for laser mode The Laser TIRF imaging system can be equipped optionally with the remote switch for laser mode which has to be connected between the Slider Laser TIRF and the distribution box for laser safety using a Y- cable.

  • Page 33: Laser Safety Concept

    INSTRUMENT DESCRIPTION Laser TIRF 3 Laser Safety Concept Carl Zeiss Laser Safety Concept The Laser TIRF imaging system is equipped with interlocks on the safety-relevant laser components. The interlocks close the laser shutter immediately if one of these components is no longer in a safe condition or is no longer in the correct work position: −...

  • Page 34: Configuration Overview Of The Laser Tirf Imaging System

    INSTRUMENT DESCRIPTION Carl Zeiss Configuration Overview of the Laser TIRF Imaging System Laser TIRF 3 Configuration Overview of the Laser TIRF Imaging System The following points should be noted when configuring a laser TIRF imaging system. • At least one option must be selected from each group (1 - 10). The components in frames are mandatory for each equipment.

  • Page 35: System Overview Laser Tirf Imaging System

    INSTRUMENT DESCRIPTION Laser TIRF 3 System Overview Laser TIRF Imaging System Carl Zeiss System Overview Laser TIRF Imaging System Fig. 2-11 System overview (page 1) 02/2014 423683-7144-001...
  • Page 36 INSTRUMENT DESCRIPTION Carl Zeiss System Overview Laser TIRF Imaging System Laser TIRF 3 Fig. 2-12 System overview (page 2) 423683-7144-001 02/2014...
  • Page 37 INSTRUMENT DESCRIPTION Laser TIRF 3 System Overview Laser TIRF Imaging System Carl Zeiss Fig. 2-13 System overview (page 3) 02/2014 423683-7144-001...
  • Page 38 INSTRUMENT DESCRIPTION Carl Zeiss System Overview Laser TIRF Imaging System Laser TIRF 3 Fig. 2-14 System overview (page 4) 423683-7144-001 02/2014...

  • Page 39: Compatibility Of Stages And Laser Safety Devices

    INSTRUMENT DESCRIPTION Laser TIRF 3 System Overview Laser TIRF Imaging System Carl Zeiss 2.9.1 Compatibility of Stages and Laser Safety Devices Incubator S TIRF S1 Incubator S TIRF DARK Laser safety Laser safety cover for (433612-9901-000) device reflected light (411857-9220-000)

  • Page 40: Compatibility Of Xlmulti Incubators And Accessories

    INSTRUMENT DESCRIPTION Carl Zeiss System Overview Laser TIRF Imaging System Laser TIRF 3 2.9.2 Compatibility of XLmulti Incubators and Accessories Inserts: Key: = compatible – = not compatible = compatible if the long camera adaptor 426118-9000-000 is used = compatible if the long camera adaptor 426118-9000-000...

  • Page 41: Overview Of Stage Versions For Tirf In Axio Observer Program

    INSTRUMENT DESCRIPTION Laser TIRF 3 System Overview Laser TIRF Imaging System Carl Zeiss Key: Inserts: = compatible – = not compatible = compatible if the long camera adaptor 426118-9000-000 is used = compatible if the long camera adaptor 426118-9000-000 is used and the CSU is mounted tilted backwards...

  • Page 42: Objectives To Be Used With The Laser Tirf Imaging System

    INSTRUMENT DESCRIPTION Carl Zeiss Objectives to Be Used with the Laser TIRF Imaging System Laser TIRF 3 2.10 Objectives to Be Used with the Laser TIRF Imaging System The functional principle described in Section 2.2 makes specific demands on the correction state of the objectives used.

  • Page 43: Fluorescence Filter Sets To Be Used With The Laser Tirf Imaging System

    INSTRUMENT DESCRIPTION Laser TIRF 3 Suitability of Fluorescence Filter Sets for the ... Carl Zeiss 2.11 Fluorescence Filter Sets to Be Used with the Laser TIRF Imaging System The following fluorescence filter sets are suitable for TIRF microscopy with Laser Bench Tower or Rack and...
  • Page 44 INSTRUMENT DESCRIPTION Carl Zeiss Suitability of Fluorescence Filter Sets for the ... Laser-TIRF 3 Wavelength Laser Filter set Typical SIRF Type Designation Order No. (nm) TIRF dyes Filter set 74 HE GFP/mRFP shift free 74HE EX DBP 480/30+565/25; 489074-0000-000 GFP/mRFP BS DFT 505+583;...
  • Page 45 = not suitable For separate observation of several dyes (double markings, etc.) please compare the corresponding filter transmissions and recommendations given on the internet at: www.micro-shop.zeiss.com | select language | Tools | Filter Assistant | Interactive Fluorescence Dye and Filter Database 02/2014...

  • Page 46: Combination Of Laser Tirf Imaging System And Colibri.2 Illumination System

    INSTRUMENT DESCRIPTION Carl Zeiss Combination of Laser TIRF Imaging System and Colibri.2 Illumination System Laser TIRF 3 2.12 Combination of Laser TIRF Imaging System and Colibri.2 Illumination System Laser Order number Beam Order number Filter set Order number Colibri.2 Colibri.2 LED...

  • Page 47: Technical Data

    INSTRUMENT DESCRIPTION Laser TIRF 3 Technical Data Carl Zeiss 2.14 Technical Data Dimensions (width x depth x height) Axio Observer stand ............... approx. 295 mm x 805 mm x 707 mm Slider Laser TIRF ..................... 280 mm x 90 mm x 80 mm Signal distribution box .................
  • Page 48 INSTRUMENT DESCRIPTION Carl Zeiss Technical Data Laser TIRF 3 Operating Data Complete TIRF system: Radio interference suppression ............in compliance with EN 55011 Class A Noise immunity ................. in compliance with DIN EN 61326-1 Laser Bench Rack / Laser Bench Rack 3F: Environment for use ......................
  • Page 49 INSTRUMENT DESCRIPTION Laser TIRF 3 Technical Data Carl Zeiss HXP 120: Environment for use ......................Closed rooms Protection class ............................I Ingress protection rating ........................IP 20 Overvoltage category ..........................II Line voltage range ..................100 V AC to 240 V AC ±10 % .....................
  • Page 50 INSTRUMENT DESCRIPTION Carl Zeiss Technical Data Laser TIRF 3 SVB 1 (signal distribution box) Line voltage ......................100 V to 240 V ±10 % Change of line voltage is not required! Line frequency ......................... 50/60 Hz, Max. power consumption ......................20 VA Fuses ..........................
  • Page 51 INSTRUMENT DESCRIPTION Laser TIRF 3 Technical Data Carl Zeiss Laser Bench Tower/Laser Bench Rack/Laser Bench Rack 3F Laser wavelength range ..................... 400 nm to 643 nm if laser protection incubator reflected / transmitted light used ............in red version..................... 400 nm to 561 nm Laser class ............................

  • Page 52: Start-Up

    The Laser TIRF imaging system has to be installed or retrofitted and adjusted in all cases by the Carl Zeiss Microscopy GmbH Service department or persons authorized by the manufacturer. If used as intended (see Section 2.1 on page 23), the Laser TIRF imaging system can be started up and used by the customer without additional precautions.

  • Page 53: Power Requirements

    START-UP Laser TIRF 3 Installation Requirements Carl Zeiss 3.1.2 Power Requirements Three wall outlets at a maximum distance of 1.5 m from the Laser Bench Tower or Rack are required to connect the system. Use the three multiple socket-outlets delivered to connect the system components (see Fig. 3-1).
  • Page 54 START-UP Carl Zeiss Installation Requirements Laser TIRF 3 Fig. 3 Terminal assignments of the multipoint connectors for the Slider Laser TIRF, Cell Observer SD (optional) and DirectFRAP (optional) combination systems 423683-7144-001 02/2014...

  • Page 55: Ambient Conditions

    Laser TIRF, is not in accordance with the intended use. In particular, Carl Zeiss Microscopy GmbH cannot guarantee laser safety in such a case. Due to the use of high-magnification objectives (e.g. α Plan-Fluar 100x/1.45 Oil), it is advisable to use a vibration protection unit Set up the microscope vibration-free on a vibration-damped table or on an anti-vibration plate.

  • Page 56: Basic Microscope Settings

    START-UP Carl Zeiss Basic Microscope Settings Laser TIRF 3 Basic Microscope Settings Be sure to read the safety data sheet provided with Immersol 518 F ® immersion oil. • Select suitable fluorescence filter sets (see Section 2.11 on page 43).

  • Page 57: Mounting The Laser Safety Cover For Reflected Light Provided For Mechanical And Scanning Stages

    START-UP Laser TIRF 3 Mounting the Laser Safety Cover for Reflected Light Provided for … Carl Zeiss Mounting the Laser Safety Cover for Reflected Light Provided for Mechanical and Scanning Stages The laser safety cover for reflected light (432319-9902-000) can be used in combination with different microscope stages.

  • Page 58: Mounting The S Tirf S1/S Tirf Dark S1 Incubator

    START-UP Carl Zeiss Mounting the S TIRF S1/S TIRF DARK S1 Incubator Laser TIRF 3 Mounting the S TIRF S1/S TIRF DARK S1 Incubator Warm air outlet Warm air inlet Locking lever for 8 Port for TIRF laser control connection cable...
  • Page 59 START-UP Laser TIRF 3 Mounting the S TIRF S1/S TIRF DARK S1 Incubator Carl Zeiss • Place the laser safety cover with the glass insert (Fig. 3-4/8) on the incubator, inserting the two metal pins into the holes on the rear side of the incubator and pressing the insert down until it slips on both sides below the front carriers.

  • Page 60: Incubator Xlmulti S1 Red Ls And Incubator Xlmulti S1 Dark Ls

    START-UP Carl Zeiss Incubator XLmulti S1 RED LS and Incubator XLmulti S1 DARK LS Laser TIRF 3 Incubator XLmulti S1 RED LS and Incubator XLmulti S1 DARK LS Handle to open the left lamp carrier sealing Left lamp carrier sealing (adjustable)
  • Page 61 START-UP Laser TIRF 3 Incubator XLmulti S1 RED LS and Incubator XLmulti S1 DARK LS Carl Zeiss The XLmulti S1 RED LS and XLmulti S1 DARK LS incubators can be used in conjunction with different stages. To check compatibility, please see the table in Section 2.9.1 on page 39.

  • Page 62: Laser Safety: Position Of The Reed Contacts

    (supporting surface for the incubator). To limit the distance of travel to the left, mount the modified strip (provided by Carl Zeiss). Before placing the incubator on the microscope, the transmitted light carrier must be moved into position, the upper illuminators removed, and the front handle (between the tube and microscope stage) removed using an Allen key.
  • Page 63 START-UP Laser TIRF 3 Incubator XLmulti S1 RED LS and Incubator XLmulti S1 DARK LS Carl Zeiss The location pins (Fig. 3-12/1 and 3) for the incubator must be screwed tightly into threaded holes microscope (Fig. 3-12/2 and 4). The pin...
  • Page 64 START-UP Carl Zeiss Incubator XLmulti S1 RED LS and Incubator XLmulti S1 DARK LS Laser TIRF 3 • Place the XL S1 heating unit to the right or left behind the incubator and connect to the incubator using the ventilation hose (Fig. 3-14).

  • Page 65: Pm S1 Ls Incubators

    START-UP Laser TIRF 3 PM S1 LS Incubators Carl Zeiss PM S1 LS Incubators The PM S1 LS incubator can be used in combination with different heating inserts and stages. The incubator can be used in combination with the following heating inserts.

  • Page 66: Mounting

    START-UP Carl Zeiss PM S1 LS Incubators Laser TIRF 3 3.7.1 Mounting The PM S1 LS incubator can be used in combination with different stages. To check compatibility, please see the table in Section 2.9.1 on page 39. Temperature sensor (inside incubator)
  • Page 67 START-UP Laser TIRF 3 PM S1 LS Incubators Carl Zeiss There are two possible ways of positioning the magnets: − (A) On the long sides of the incubator, in extended lines to the red markings. − (B) On the short sides of the incubator. In this case, ensure that the middle of each magnet (Fig.

  • Page 68: Assembly Of The Magnet Screw

    START-UP Carl Zeiss PM S1 LS Incubators Laser TIRF 3 3.7.2 Assembly of the Magnet Screw The magnet screw consists of three parts. • First screw the M5x12 (Fig. 3-18/3) screw into one of the threaded holes (Fig. 3-18/4) on the microscope stand close to the transmitted light illumination carrier using an Allen key (SW 4).

  • Page 69: S Tirf S1 And S Tirf Dark S1 Incubators

    START-UP Laser TIRF 3 S TIRF S1 and S TIRF DARK S1 Incubators Carl Zeiss S TIRF S1 and S TIRF DARK S1 Incubators The S TIRF S1 and S TIRF DARK S1 incubators can be used in combination with different stages. To check compatibility, please see the table in Section 2.9.1 on page 39.

  • Page 70: Insert Slider Laser Tirf

    START-UP Carl Zeiss Insert Slider Laser TIRF Laser TIRF 3 Insert Slider Laser TIRF • If necessary, remove the luminous-field diaphragm slider from the stand. • Carefully push the Slider Laser TIRF into the receptacle of the luminous-field diaphragm of the microscope until it clicks into place (Fig.

  • Page 71: Operation

    OPERATION Laser TIRF 3 Control and Functional Elements Carl Zeiss OPERATION The Laser TIRF imaging system is a class 3B laser system and labeled accordingly. CAUTION: This risk category includes medium-power lasers. Ensure that you are not exposed to the radiation of such lasers. In particular never look into the laser beam! Control and Functional Elements 4.1.1...

  • Page 72: Laser Bench Tower

    OPERATION Carl Zeiss Control and Functional Elements Laser TIRF 3 The Active LED (Fig. 4-1/4) lights up when laser operation mode is activated. Laser operation mode can be activated/deactivated either via the button ring of the microscope, using the software, or the remote switch for laser mode.

  • Page 73: Laser Bench Rack

    OPERATION Laser TIRF 3 Control and Functional Elements Carl Zeiss 4.1.3 Laser Bench Rack Key switch (Laser ON / OFF) Fig. 4-3 Control and functional elements of the Laser Bench Rack 02/2014 423683-7144-001...

  • Page 74: Switching On

    The device is ready for use when the Active and Safe LEDs are lit. See Sections 4.4 to 4.7. If the safety system does not work in the way described above, take the instrument out of use, label it clearly as being out of order and immediately contact Carl Zeiss Microscopy GmbH Service.

  • Page 75: Switching Off

    The Slider Laser TIRF mot. is controlled via the ZEN software (blue edition) using the Calibration Wizard. Calibration of the Slider Laser TIRF mot. via software requires the camera adapter 60N-C 1.0x for TIRF 3 with Bertrand optics. 4.5.1 Delivery Scope of the TIRF Adjustment Tool Kit The following components are included in the TIRF adjustment tool kit (423684-9010-000) delivered: −...
  • Page 76 OPERATION Carl Zeiss Checking and Adjusting the Laser Beam Divergence and the TIRF Angle Laser TIRF 3 − 2x Adapters M27x0.75 to W0.8; H=2.56 mm, − 2x Adapters M27x0.75 to W0.8; H=4.24 mm, − 2x Adapters M27x0.75 to W0.8; H=6.85 mm, −...

  • Page 77: Adjusting The Laser Beam Divergence Using The Adjustment Tool

    OPERATION Laser TIRF 3 Checking and Adjusting the Laser Beam Divergence and the TIRF Angle Carl Zeiss 4.5.2 Adjusting Laser Beam Far out of focus Divergence Using the Adjustment Tool Huge diameter 1. Focus on the glass/medium interface of the specimen (adherent beads) in epi-fluorescence.

  • Page 78: Adjusting And Checking The Tirf Angle Using The Adjustment Tool

    OPERATION Carl Zeiss Checking and Adjusting the Laser Beam Divergence and the TIRF Angle Laser TIRF 3 4.5.3 Adjusting and Checking the TIRF Angle Using the Adjustment Tool 1. Focus on the glass/medium interface of the specimen (adherent beads) in epi-fluorescence.

  • Page 79: Adjusting The Tirf Effect On The Tirf Slider Man

    OPERATION Laser TIRF 3 Adjusting the TIRF Effect on the TIRF Slider man. Carl Zeiss Adjusting the TIRF Effect on the TIRF Slider man. • Place a specimen in the object mounting frame. • Move the required TIRF objective into the optical path.
  • Page 80 OPERATION Carl Zeiss Adjusting the TIRF Effect on the TIRF Slider man. Laser TIRF 3 • Now activate epi-fluorescence illumination by pressing the RL button on the right-hand side of the microscope stand. • Set the beam path of the microscope in such a way that the light from the specimen is deflected to the camera.

  • Page 81: Controlling The Laser Tirf Imaging System Using Zen 2012 (Blue Edition)

    OPERATION Laser TIRF 3 Controlling the Laser TIRF Imaging System Using ZEN 2012 (Blue Edition) Carl Zeiss Controlling the Laser TIRF Imaging System Using ZEN 2012 (Blue Edition) The basis of the following outline of how to integrate the TIRF imaging system laser into the ZEN 2012 software (blue edition) is the corresponding software manual or the online help function of the program.

  • Page 82: Calibration Of The Angle Using The Motorized Slider Laser Tirf

    OPERATION Carl Zeiss Calibration of the Angle Using the Motorized Slider Laser TIRF Laser TIRF 3 Calibration of the Angle Using the Motorized Slider Laser TIRF 4.8.1 Measurement of Penetration Depth The penetration depth of the evanescent field under TIRF conditions is determined using the following equation.

  • Page 83: The Angle-Voltage Relationship Of The Slider Laser Tirf

    OPERATION Laser TIRF 3 Calibration of the Angle Using the Motorized Slider Laser TIRF Carl Zeiss 4.8.3 The Angle-Voltage Relationship of the Slider Laser TIRF The following correlation applies for the deflection of the laser beam in the rear objective pupil: U −...

  • Page 84: Determining The Critical Angle

    OPERATION Carl Zeiss Calibration of the Angle Using the Motorized Slider Laser TIRF Laser TIRF 3 4.8.4 Determining the Critical Angle The offset of the deflection can be determined using the actual sample. The angle is moved into the TIRF position on both sides of the pupil using a software calibration routine.

  • Page 85: Care, Maintenance, Troubleshooting And Service

    CARE, MAINTENANCE, TROUBLESHOOTING AND SERVICE Laser TIRF 3 Instrument Care Carl Zeiss CARE, MAINTENANCE, TROUBLESHOOTING AND SERVICE Instrument Care Care of the instrument by the user is restricted to the following operations: The instruments are not equipped with any special devices for protection against substances that are corrosive, potentially infectious, toxic or radioactive, or other substances that could be hazardous to health.

  • Page 86: Troubleshooting

    Do not remove the connections from the Laser Bench Tower or Rack under any circumstances. The Laser Bench Tower and Rack may only be connected by Carl Zeiss Microscopy GmbH Service staff. • Check that the connections on the Laser Bench Tower and Rack are tight.

  • Page 87: Fuse Replacement

    Make sure the power cable is disconnected before changing any fuses. Axio Observer, HBO, XBO, HXP, Tempmodul S1: see separate operating manual. Laser Bench Rack and TIRF Controller: Do not contain fuses changeable from outside. Please contact the Service Department of Carl Zeiss Microscopy GmbH. 5.4.1 Signal Distribution Box The fuse compartment is combined with the coupler plug and contains two fuses.

  • Page 88: Laser Bench Tower Coupling Module

    The laser must always be switched off before changing the halogen lamp of the HAL illuminator! See separate operating manual. Retrofitting the Laser Units The laser units may be retrofitted by Carl Zeiss Microscopy GmbH only! Retrofitting will be charged additionally. 423683-7144-001 02/2014...

  • Page 89: Mounting The Camera

    CARE, MAINTENANCE, TROUBLESHOOTING AND SERVICE Laser TIRF 3 Mounting the Camera Carl Zeiss Mounting the Camera The camera is connected to the sideport of the microscope using the camera adapter. To ensure safe operation of the laser, the camera adapter is fitted with two reflected light barriers which close the laser shutter when no camera (Fig.

  • Page 90: Connecting The Hxp 120 Illumination Unit

    CARE, MAINTENANCE, TROUBLESHOOTING AND SERVICE Carl Zeiss Connecting the HXP 120 Illumination Unit Laser TIRF 3 Connecting the HXP 120 Illumination Unit The HXP 120 illumination unit is connected by means of a coupling adapter, in which the optical waveguide is firmly secured and aligned, to the reflected light illumination connector of the microscope.

  • Page 91: Service Request

    To ensure optimum settings and trouble-free functioning of your Laser TIRF imaging system over a longer period of time, we recommend you conclude a service / maintenance agreement with Carl Zeiss Microscopy GmbH. For subsequent orders or service requests, please contact the Carl Zeiss Microscopy GmbH representative responsible for you.

  • Page 92: Using The Slider Laser Tirf With Customers' Own Lasers

    Service Request Laser TIRF 3 USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS The following pages do not apply for TIRF systems which have been set up by Carl Zeiss Service staff. Advice for users with their own laser modules: Only use class 3B laser modules (maximum 500 mW of total output power).

  • Page 93: Laser Safety Concept

    USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Laser TIRF 3 Laser Safety Concept Carl Zeiss Laser Safety Concept The Laser TIRF imaging system is equipped with interlocks on the safety-relevant laser components. The interlocks close the laser shutter immediately if one of these components is no longer in a safe condition or is no longer in the correct work position: −...
  • Page 94 USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Carl Zeiss Laser Safety Concept Laser TIRF 3 Fig. 6-2 Connection overview (schematic) 423683-7144-001 02/2014...

  • Page 95: Safety Kit For Customer-Provided Laser Module

    USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Laser TIRF 3 Safety Kit for Customer-Provided Laser Module Carl Zeiss Safety Kit for Customer-Provided Laser Module The Safety Kit TIRF for customer-provided laser modules (432921-0000-000) includes a safety shutter for the laser module (laser shutter) and a connection cable.

  • Page 96: Remote Switch For Laser Mode

    USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Carl Zeiss Remote switch for laser mode Laser TIRF 3 Remote switch for laser mode The Laser TIRF imaging system can be equipped optionally with the remote switch for laser mode which has to be connected between the Slider Laser TIRF and the distribution box for laser safety using a Y- cable.

  • Page 97: Connecting The Remote Switch For Laser Mode

    USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Laser TIRF 3 Connecting the Remote Switch for Laser Mode Carl Zeiss Connecting the Remote Switch for Laser Mode • Connect one connector of the Y-cable to the safety cable of the Slider Laser TIRF (Fig. 6-5/2).

  • Page 98: Basic Kit Laser Safety Rev. 2

    USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Carl Zeiss Basic kit laser safety Rev. 2 Laser TIRF 3 Basic kit laser safety Rev. 2 The basic kit laser safety Rev. 2 includes an eyepiece shutter (factory-installed or retrofitted by service personnel), a laser safety board for the stand, a distribution box and sticker labels.
  • Page 99 USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Laser TIRF 3 Basic kit laser safety Rev. 2 Carl Zeiss Connection plug laser safety cover / incubator Connection plug laser shutter for customer's own laser module Connection plug Slider Laser TIRF...
  • Page 100 Carl Zeiss Basic kit laser safety Rev. 2 Laser TIRF 3 • Connect the 25-pin sub-D connector (Fig. 6-7/5) of the distribution box for laser safety (Fig. 6-7/6) to the jack for Laser TIRF on the rear of the microscope stand (Fig. 6-7/10).

  • Page 101: Connecting The Slider Laser Tirf Mot

    Switch the laser off before mounting the optical fiber! If customers use their own laser, the Carl Zeiss Monomode Fiber 400615-9400-000 may not be used. 02/2014...
  • Page 102 USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Carl Zeiss Coupling a Laser to the Laser TIRF Imaging System Laser TIRF 3 • Fig. 6-9/A: Remove the two Allen screws (Fig. 6-9/2) and push the unscrew protection cap (Fig. 6-9/3) over the optical fiber to be connected (Fig.

  • Page 103: Safety Check Prior To First Start-Up

    USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Laser TIRF 3 Safety Check Prior to First Start-up Carl Zeiss Safety Check Prior to First Start-up Prior to the first start-up and after every re-installation of the Laser TIRF imaging system, please check the laser safety system for its ability to operate properly.
  • Page 104 If the safety system does not work in the way described above, take the instrument out of use, label it clearly as being out of order and immediately contact Carl Zeiss Microscopy GmbH Service. Attach the microscope tube again after successful safety check.

  • Page 105: Power Requirements

    USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Laser TIRF 3 Power Requirements Carl Zeiss Power Requirements Three wall outlets at a maximum distance of 1.5 m from the Laser Bench Tower or Rack are required to connect the system.
  • Page 106 USING THE SLIDER LASER TIRF WITH CUSTOMERS' OWN LASERS Carl Zeiss Power Requirements Laser TIRF 3 Fig. 6 Terminal assignments of the multipoint connectors for the Slider Laser TIRF, Cell Observer SD (optional) and DirectFRAP (optional) combination systems 423683-7144-001 02/2014...

  • Page 107: Appendix

    APPENDIX Laser TIRF 3 List of Abbreviations Carl Zeiss APPENDIX List of Abbreviations AOTF Acousto-Optical Tunable Filter Canadian Standards Association Deutsches Institut für Normung (German Standards Institute) European standard Electromagnetic Compatibility Mercury-vapor short-arc lamp for fluorescence Mercury short-arc lamp International Electrotechnical Commission...

  • Page 108: Index

    APPENDIX Carl Zeiss Index Laser TIRF 3 Index Page Ambient conditions ..........................48 Basic laser safety kit ........................31, 99 Basic settings ............................57 Care ..............................86 Checking the system ...........................86 Configuration .............................34 Connect distribution box for laser safety ..................87, 100 Connecting HXP ..........................91 Control and functional elements ......................72...
  • Page 109 APPENDIX Laser TIRF 3 Index Carl Zeiss Maintenance ............................86 Mounting the camera ......................... 90 Mounting the incubator ........................59 Mounting the laser safety cover ......................58 Notes on warranty ..........................22 Objectives............................43 Operating principle ..........................24 Operation ............................. 48, 72 Optical fiber ............................

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