Инструкция для OLYMPUS CELL FRAP

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 Manual

 

Chapter 6 – 

Operating the 

 

System

 

35 

 

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 happen?

 

 

After  the  start  of  the  experiment  the  FRAP  command  icon  will  simply  be  ignored  until  clicked  by  the 

user.  Clicking  is  possible  at  any  time  and  will  immediately  start  the  ROI  scan  independently  of  the 

position of the command icon in the experiment plan and independently of the current advance of the 

experiment. 

Preview 

 

Live view while bleaching 

button 

Click  here  to  start  a  live  preview  simultaneously  with  a  laser  scanning  cycle.  The  scanning  laser  spot 

will be visible in the beginning. 

 

Live view after bleaching 

button 

Click  here  to  start  a  laser  scanning  cycle  with  a  live  preview  following  immediately  after  the  scan  is 

finished. 

 

Snapshot after bleaching 

button 

Click here to start a laser scanning cycle with a single snapshot following immediately after the scan is 

finished. 

 

Stop FRAP preview 

button 

Click here to stop the 

Live view

Extended 

Scan  speed

.  This  determines  the  duration  of  a  scan  and  thus  the  time  that  the  laser  rests  at  each 

position  during  the  scan.  Set  the  scan  speed  by  use  of  the  slider  or  by  typing  a  value.  The  speed  is 

expressed as frequency in Hz. The default value is 10 kHz (10.000 Hz). 

FRAP inside ROI

. Mind that the ROIs drawn in the image will be "filled" with positions of the laser spot 

and thus the form of the ROIs will only be approximated. This parameter defines how much the outer 

laser  spot  positions  may  overlap  with  the  ROI  borders.  Set  the  percentage  by  use  of  the  slider  or  by 

typing a value. The default value is 100%. 

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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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