Инструкция для OLYMPUS CELL FRAP

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 Manual

 

Chapter 6 – 

Operating the 

 

System

 

31 

 

8.

 

TTL Synchronization

FRAP Trigger OUT -> DIG I/O

. Select from the short list the TTL IN 

connection  at  the 

  imaging  PC  front  panel  that  is  connected  with  the 

TRIGGER 

OUT 

of the 

 control unit. 

9.

 

Exit the software with 

OK

6.2

 

The FRAP GUI in 

 

This  interface  is  mainly  meant  for  calibration  purpose  and  for  opening  and  closing  the  FRAP  laser 

shutter. The use of 

 in the context of "true" FRAP experiments is explained in the next Chapter 

6.3, 

 

in the Experiment Manage

r. 

 

Control 

Disconnected  /  Connected

.  The  box  turns  green  and  reads 

Connected

  if  the  software  detects  the 

presence  of  the 

  controller  via  the  serial  connection.  The  status  can  be  update  by  clicking  the 

Update

 button on the right side. 

Laser  on/off

.  Use  this  toggle  button  to  turn  the  laser  connected  to  the 

  (and  configured 

accordingly in OBS System Configuration) on or off (respectively, open or close its shutter). 

Center  spot

.  Click  this  button  to  move  the  scanner  mirrors  to  their  central  position.  This  will  set  the 

laser spot to the central image position if the system is aligned correctly. 

Objective

. This field displays the currently active objective. The calibration needs to be performed for 

each objective that is to be used for the FRAP experiments. The currently used objective is listed here. 

Calibrated

. The status box turns green once the system is calibrated. 

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Оглавление инструкции
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    ® I INSTRUCTIONS SOFTWARE MODULE AND LASER SAFETY FOR THE HIGH-SPEED SCANNING FRAP SYSTEM This instruction manual describes the configuration and usage of the scanning FRAP Imaging Stations. To ensure safety, obtain optimum system within the Olympus performance and familiarize yourself fully with
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    OLYMPUS SOFT IMAGING SOLUTIONS GMBH Johann-Krane-Weg 39 D-48149 Münster Tel: +49 89 - 89 55 805 660 Fax +49 89 - 89 55 805 6606 Email: info@olympus-sis.com www.olympus-sis.com
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    III Imaging Excellence We at Olympus Soft Imaging Solutions GmbH have tried to make the information in this manual as accurate and reliable as possible. Nevertheless, Olympus Soft Imaging Solutions GmbH disclaims any warranty of any kind, whether expressed or implied, as to any matter whatsoever
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    Manual Contents Contents 1 Introduction.........................................................................................................1 1.1 Abstract ........................................................................................................2 1.2 General
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    Contents
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    Manual Chapter 1 –Introduction 1 Introduction Thank you very much for purchasing Olympus Soft Imaging Solutions' state of art scanning FRAP system and for your confidence in our products and service. It is Olympus Soft Imaging Solutions' main objective to provide you with solutions able to meet
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    2 Chapter 1 – Introduction scan head mounted on an IX81 1.1 Abstract In the following sections you will find a detailed description of the components of scanning FRAP system, handling and alignment instructions and how to maintain the performance of your FRAP microscope. Keep in mind that all
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    Manual Chapter 1 –Introduction 1.2 General The MITICO motorized integrated TIRF illumination combiner is compatible with the Olympus microscope frames IX81S1F-FRAP, IX81S1F-FRAP-ZDC, and IX71S1F-FRAP, as well as a range of diode laser systems between 405 nm and 700 nm provided that they meet the
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    4 Chapter 1 – Introduction a illuminator featuring two fiber ports, each with adjustable line focus b control unit
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    Manual Chapter 2 – Safety Symbols 2 Safety Symbols The deactivation of the safety systems or the dismantling of different components by the operator is required in certain operational modes and in case of illumination alignment. As a result the Olympus scanning FRAP system is to be classified in
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    6 Chapter 2 – Safety Symbols The following symbols are placed on the Olympus scanning FRAP system. Study the meaning of the symbols and always use the equipment in the safest possible manner. General symbols Symbol Explanation Indicates the use of a laser beam. Take special care in handling that
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    Chapter 2 – Safety Symbols Manual 1: scan head 2: microscope frame, right side 3: fluorescence turret and C-mount 4: laser safety ocular shutter 5: laser safety stage cover 7
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    8 Chapter 2 – Safety Symbols
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    Illumination Combiner, User Manual Chapter 3 – Safety Precautions and Handling Instructions 3 Safety Precautions and Handling Instructions The Olympus scanning FRAP system must be used with laser products belonging to the Laser Safety Class 3B. Class 3B lasers are medium power lasers with an output
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    10 Chapter 3 – Safety Precautions and Handling Instructions 3.1 Laser Safety Precautions Caution: Control or adjustments using procedures other than those prescribed in this manual will lead to hazardous laser beam exposure. To avoid hazardous situations strictly follow the laser safety
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    Illumination Combiner, User Manual Chapter 3 – Safety Precautions and Handling Instructions Direct exposure to the laser light is possible at the following positions: • The laser system’s housing at the fiber port if the fiber is removed • The exit of the laser fiber • The fiber coupling of the
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    12 Chapter 3 – Safety Precautions and Handling Instructions
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    Manual 4 Chapter 4 – Overview and Laser Safety Setup Overview and Laser Safety Setup The following chapter describes all components and interfaces of the Olympus system including all laser safety devices and how they are installed and connected. 4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.8 scanning FRAP Setup
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    14 Overview and Laser Safety Setup Chapter 4 – 4.1 Setup Scheme Laser fibers LAS-BOB scan head Laser safety stage cover FRAP lasers with safety shutters Laser safety ocular shutter LAS-MC IX81, top view controller Scheme of an IX81 microscope with a PC scanning FRAP system, two lasers and the laser
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    Manual Chapter 4 – Overview and Laser Safety Setup 4.2 The Laser Safety Features – Overview and Check The laser safety concept of the scanning FRAP system consists of the following modules: – A laser safety ocular shutter – A laser safety stage cover – An emergency interrupter located on the LAS-MC
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    16 Chapter 4 – Overview and Laser Safety Setup Laser safety stage cover Laser safety stage cover • The 5-pin ocular shutter cable must be connected to the stage cover and this additionally to one of the 5-pin SAFETY INTERLOCK plugs at the back panel of the control unit. • The other 5-pin SAFETY
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    Manual Chapter 4 – Overview and Laser Safety Setup 4.3 Mounting the Laser Safety Kit 1. Remove the ocular from the microscope frame and mount the ocular laser safety shutter onto the ocular tube (with the cable on the right hand side) and fix it as shown in the photo below. 2. Mount the ocular back
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    18 Chapter 4 – Overview and Laser Safety Setup 6. Connect the second plug of the laser safety stage cover with the 5-pin female STAGE COVER plug of the scanning illuminator using the corresponding cable. Translucent insert 4.4 Connecting the 5-pin plugs Control Unit The installation of the scanning
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    Manual Chapter 4 – 3. Connect the PC RS232 plug at the back panel of the the system PC. Overview and Laser Safety Setup control unit with a serial port of 4. Connect the cable coming from the scan head with the multi-pin plug at the front panel of the control unit. 5. Connect the TRIGGER IN TTL BNC
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    20 Chapter 4 – Overview and Laser Safety Setup 2. Connect the 5-pin female STAGE COVER plug at the LAS-BOB back panel with one of the control unit using 5-pin female SAFETY INTERLOCK plugs at the back panel of the the corresponding cable. Make sure the control unit is connected with the laser
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    Manual Chapter 4 – Overview and Laser Safety Setup – A mode lock with key to allow authorized and trained personnel to switch from standard USER MODE to MAINTENANCE (see Chapter 5, Modes of Operation). – A red emergency interrupter that closes all safety shutters when pressed. – A BINOCULAR button
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    22 Chapter 4 – Overview and Laser Safety Setup 4.8 The Remote Interlock Connector of the Olympus Soft Imaging Solutions ODB Laser Systems An INTERLOCK connector can be found at the front panel of all Olympus Soft Imaging Solutions laser systems as Class 3B laser products. This remote interlock
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    Manual Chapter 5 – Modes of Operation 5 Modes of Operation There are two modes of operation, the user mode and the maintenance mode. In user mode no laser radiation can exit the system. The maintenance mode is for setting-up and adjusting the scanning FRAP system. Only trained personnel are allowed
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    24 Chapter 5 – Modes of Operation 5.1 The User Mode All hardware has to be connected properly to operate the system in User mode. The user mode ensures that the user can operate the system without being exposed to the laser beam. It is possible to have full control of the various devices via
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    Manual Chapter 5 – Modes of Operation To protect your specimen from photo-bleaching, close the laser shutter (by reversing step 5) whenever suitable, even if you interrupt observation only for a short time. 5.2 The Maintenance Mode Actions in maintenance mode Permitted Action Remarks Exchanging
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    26 Chapter 5 – Modes of Operation 5.2.2 Access to the Specimen without Deactivating the FRAP System For some applications it is necessary to manipulate the specimen during an experiment. In User mode, however, the laser safety stage cover has to be in place; otherwise the FRAP system is deactivated
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    Manual Chapter 5 – Modes of Operation Laser safety ocular shutter, slider OUT 3. Hold a sheet of paper in front of the eyepieces. 4. Open the laser safety shutters of the lasers by pressing the red ocular safety interlock bridge button of the LAS-MC manual control. Check the sheet of paper to
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    28 Chapter 5 – Modes of Operation
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    Manual Chapter 6 – Operating the 6 Operating the System System The scanning FRAP system and the Olympus Soft Imaging Solutions laser systems are fully integrated in the imaging software This chapter describes the configuration and usage of the scanning FRAP system. 6.1 6.2 6.3 6.3.1 6.3.2 6.3.3
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    30 Chapter 6 – Operating the System 6.1 System Configuration 1. Start the ObsConfig software to configure the excitation filters 2. Make sure that the FRAP lasers are configured on pages Microscope Laser as described in the laser manual. 3. Go to page Microscope FRAP. In case this page is not
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    Manual Chapter 6 – Operating the System 8. TTL Synchronization, FRAP Trigger OUT -> DIG I/O. Select from the short list the TTL IN imaging PC front panel that is connected with the TRIGGER connection at the OUT of the control unit. 9. Exit the software with OK. 6.2 The FRAP GUI in This interface is
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    32 Chapter 6 – Operating the System The calibration is valid only for the objective in place during the calibration. A later manual change of the objective does not cause a change of the status display. However, a recalibration is required. Set spot size. Click here to open the Enter spotsize
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    Manual Chapter 6 – Operating the System because the software will detect the bright spot when it is reasonably close. Increase the step size by keeping the Ctrl key pressed while using the arrow key. Keep the Shift key pressed to reduce it. Once the spot is at the position of the red circle click
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    34 Chapter 6 – Operating the System after bleaching. The Acquire radio buttons and the Condition parameters are deactivated. Manual start during experiment is not possible. FLIP (Fluorescence Loss In Photobleaching). In this experiment the ROIs are bleached in Every Cycle and acquisition always
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    Manual Chapter 6 – Operating the System Manual start during experiment. This check box is active only if the Advanced experiment is selected. If the option is checked the command icon shows a hand symbol. The command icon turns into a button. FRAP command icon with Manual start symbol What will
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    36 Chapter 6 – Operating the System Edge accuracy. The FRAP scanner controller composes the ROIs to be scanned out of subrectangles. The smaller the sub-rectangles are the higher is the resulting correspondence with the ROIs. Set the accuracy by use of the slider or by typing a value. The default
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    Manual Chapter 6 – Operating the System is the first one within each cycle. At one point in time during a certain cycle a Click is being applied. The Fire will only be carried out as soon as the next cycle is being started. Thus, while the manual Click is unsynchronized, of course, the Fire
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    38 Chapter 6 – Operating the System
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    Manual Chapter 7 – Specifications, Technical data 7 Appendix This chapter lists specifications and technical data of the 7.1 system. Laser Safety Stage Cover Dimensions .......................................................40 39
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    40 Chapter 7 –Specifications, Technical Data 7.1 Laser Safety Stage Cover Dimensions The laser safety stage cover must be secured to the microscope stage with the M4x5 screws provided. If the microscope stage does not have the necessary pre-drilled holes, this simple modification must be performed
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    Manual Chapter 7 – Specifications, Technical data 41
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    ® OLYMPUS EUROPA HOLDING GMBH Postfach 10 49 08, 20034, Hamburg, Germany Printed in Germany 2010 09 M 001
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