User guide spinning disk
Contents
1. 4 AOTF Shutter Open Closed a a saree Click Next three times until you arrive at the AOTF settings for the Andor Laser Combiner Power mW max Laser Status FRAPPA 15 0 250 0 _sewok__ f Here you can modify the laser power for the FRAP Ensure the laser Laser Line AOTF T z os E a7 you want to FRAP with is ticked i a si im Fae so r E T o M Shutter NB Make sure FRAPPA is selected in the menu at the bottom If it isn t your FRAP won t work MPU OESE FRAPPA Once this is completed repeat the test FRAP If its ok you can run the FRAP protocol as you would any other protocol i Run 12 TIRF e e Acquiring multiple positions e g in a multiwell plate can be xy or xyz s PS Andor iQ 2 6 Andor Camera olx L Make sure you have registered the stage if not Fie Image List Settings Wizards Process 4nalysis Plug Ins Help lower the objective so it doesn t crash into the 5 ETE 2 a j oe stage Go to Wizards in the menu and choose Save Idle Snap CSU 5B8 Quad Stage Registration 2 Select a Protocol with a xy scan in it Protoce i Protocol 3 Select the Repeat XY in the protocol and choose Edit Fee The XY wizard will ask if you want to move the Ea 985_10px01 0 06667 stage to the stored position Select No Ey Protocol ET scar Protoco Sgad Image Red imaging Ea Protocol otherwise the stage will move so2. Series12 This has a list that has been autosaved in the computer s Pah inapeDok Or memory of all the experiments you have done Please select fet 0 80000 these and delete the experiments carried out during your imaging session from the image list Uses M aba NO INE INN Image List NB If the image list gets full the machine will crash so this is important E 2 If you have accidentally forgotten to save an experiment you can find it in the image list and click show This will reload your experiment You will then be able to save it b IME 2 avele Time 2 i 03 057 S eies 10 227 03 057 14 bit gr oF Wavele Time 2 Close Refresh Show Rename Image Save Image Delete Images To make a Z stack The z scan software works by selecting the top and bottom of the sample For very precise mechanical scanning the Piezo drive is used So you need to navigate through the stack using the Piezo motor DO NOT TOUCH THE FOCUS ON THE MICROSCOPE STAND 1 Open the protocols menu as for Time series m tata ID User Data Autol mages 2 2 Select Z Stack from the options Acquisition Calibration poe 885_100Xoi 0 06667 Elf Protocol Z stack Protacal St m Image Z stack ue ait Camera Binning Use Current les Protocol Wait Allocation Off all Camera Selection Use Current 2 Run ii Z Channel In the Z protocol select Repeat Z and choose Edi
3. e The FRAPPA bleaches the sample by point scanning It will take a short time 1 sec for the system to switch back to spinning disk confocal imaging please allow for this in your experiment e For good recovery you need to bleach your dye by about 50 e The camera noise level is set to read an intensity of around 300 In order to quantify FRAP you will need the structure you are hoping to FRAP to have an intensity of 1000 or more e The FRAP doesn t bleach a point It bleaches the point spread function in your image i e the focal planes above and below where you bleach will also be bleached e FRAPPing with too much laser power may kill your cell 1 Snap an image of the sample you want to FRAP 2 Select FRAPPA from the ROI menu PS RODS 6 TIF 14 bit 1004 0 067 x 1002 0 067 3D File Edit Image List Region View Fast MIP Tate ala Nile Clojols Ti meN Background Crop Edit Classifications TO JO s TI ima eo Draw the ROI you are interested in on your image NB no more than 8 sides Select the FRAPPA Channel by right clicking in the image Select Toal wf Auto Map Select the FRAP laser you want Tick appears Copy next to laser Faste Edit Delete Delete All To test FRAP click FRAPPA this Classify Load Save Mave To FRAPPA 405 FRAFFA 561 frappa 647 Set Reference Frappa This Frappa On Mouse Up Frappa Device Frappa Chann
4. 000 E I Select All Gi version Info ate Stop All Ar 15 The start up axes menu will appear If Autozero says Not Ok the device needs to be auto Zeroed Start up stages axes for E 753 on RS 232 COM7 9600 baud Select all Select unreferenced Select axes with stored states Ref switch Limit hard s liz P 25 1C0 no on target 79 398800 OK 2 ect TEA ages Eaa E anaes ae Start up axes Reference Mlected axes by moving to Reh switch Auto zera Automatic Help Close Once the device is Auto zeroed the Z position which the PiFoc will maintain needs to be set To do this got to Tools and select Single axis window and select the PiFoc the only option pPIMikroMove 2 9 0 4 E a iolz Connections E 753 COMA Tools view Help a a ee OY afa ais D Single Axis Window 1 2 P 725 1CD on E 753 on RS 232 COM 9600 baud f ro E Bigitel oa Window a Move multiple axes Er E s SS l Host macros C Piezo channels Elsensor channels Axis Target IY 1 2 P 725 1CD 80 E Stage l g Openloop Value gt Step size Position HALT State elocity Servo m 1 z oFIP F25 1cD 80 000 76 846 gt 0 100 79 995 BRS on target 10000 000 w ie I Select All P Use vector move where possible i E 2 pa ir At Stop All Start E The c
5. 6 Settings Exposure General Device Info Status a4 Window Size 1004 1002 Gai Full Chip Window fw EM Gain Enabled 297 Getting Started First of all select the protocol for your experiment The easiest way is to choose Protocol from the menu in the left hand side dialogue You can also go to protocols Using Wizards and Protocols A list of protocols will appear Pick the one you want and press Select Andc iQ 1 10 2 Andor Camera fx File ImaMeList Settings QC Wizards Process Analysis Plugins Hg Setup Vizard Protoc ls Protokol Manager E og Select Cancel New Delete Copy Protocol ummaries Currer Protocol pcan 4 Protocol Frap 4 Wrotocol Time series and FRAP 4 Protocol Time series 4 Protocol z stack The protocol for the imaging experiment you are carrying out will then appear in the left hand side dialogue box Andor iW oD Ano LaMmera ra E Jy xy File Image List Settings Wizards Process Analysis Plug Ins Help cs U 568 0 uad Acquisition Analysis AGB Analysis Movie Editor Spot Analysis Status eT Auto Saver E User Data Aautolmages i Acquisition Calibram sel See AAOXoi 0 06667 El Protocol Two channel Timelapse Protoca cgn Image Two channel T imelapse ui Camera Binning Use Current a walt Allocation Of von Camera Select
6. 7 Frame Averaging 2 il fioo578 fms z Poema 2 Ef Protocol Two channel Timelapse Protocot RET EA B em Image Two channel Timelapse a Readaik ms 32 723 a toe ast Refresh a Camera Binning Use Current eA f Fast Retr p Wait Allocation Off O Bun Display Every Frame Frame Rate Camera Selection Use Current C Display 1 Frames s ls Actual Rate 0 00 H S Events Q Chamel B Q Repeat T 60 times 5 sec P hannal m e Move Channel CSU 488 Quad XY Scan Snap Frame m e Move Channel CSU 568 Quad J Device Setup Trigger Controt Snap Frame Intemal Trigger Ext Trigger Timeout ms foo Loop Protocol End 1 3 ce ioe 7 ExtemalStat PC RAM MB 2047 IV Show All M Camera Status Camera is working te SE Repeats 60 Save As L ames ike Bind 4 Interval B sec M a Options Cancel 2 Help Piezo channels E Sensor channels arget Openloop Yalue gt gt Step size Position HALT State Yel 60 000 6 024 gt gt 0 100 80 000 IA ontarget 10i Time Wavelength 0 02 00 009 A him s ms Frame Number Minrange Maxrange Increment a Interval s ms a af gt 24 fo 24 fi oor a alial m fom ri ala StM Set Max Flatten Time ez az Pam os reas ee E This number tells you the intensity of the pixel your mo
7. OPERATING INSTRUCTIONS Spinning disk You must not operate this equipment without prior training from a BALM facility staff member To arrange training and for help please contact Facility Manager Dr Ann Wheeler ext 2406 a p wheeler qmul ac uk Microscopy Technologist Isma Ali ext 2407 i ali qmul ac uk Standard Operating Procedure How to turn the equipment on Switch on the three switches A Mercury halide lamp B and C left hand side on wall Switch on the computer and login Wait 5 minutes for all of the equipment to initialise Clean the objective lens you want to use Open the software by hitting Andor Technology icon How to turn the equipment off Follow the instructions beside the microscope to switch off the argon laser allowing time for it to cool the computer the microscope and the metal halide lamp Rules of use This microscope should be treated with respect and care at all times The SOP must be followed This Microscope can only be used by Masters by Research or PhD students Postdocs and members of staff The microscope lenses must be cleaned after every usage and the equipment treated carefully at all times If you have any problems at all with the microscope no matter how trivial they may seem please see a member of facility staff immediately e Please ensure all sharps and clinical waste are disposed of in the sharps bin and clinical waste bin provided REMEMBER Check before you start if you
8. d 3 Focus using the Z position menu 4 Press Next Field and repeat for the next position Next Field gt gt f Position lt lt Eig 49 99 I 2552553 13 Acquisition Lalibratian 665_100 01 0 06667 Once all the positions for all fields are set the menu will go back to the protocol Protoca Protocol E Pot Protocol 7 scan E ga Image Red imaging ony A Camera Binning Use Current ao Wait Allocation Off E Camera Selection Use Current Q Pun Select the time repeat for how oof Scan test 3 be eee 8S Channel many times you would like each le Events ee pRepeat T 5 times 1 min MEY Sean position to be acquired Right click Ele Repeat sr Hl XY Positions 5 Ha Move Channel CSU 568 Guad ie Snap Frame HA Move Channel CSU 488 Guad on Repeat in the protocol oe Device Setup Loop Protocol M Display Image M Show Al Save As at Options Remember that the stage will need to move and the system will need to Frame average when working out your timelapse interval Edit repeat s E 7 Hel hp Repeats B Interval __ a Once this is complete you can run your protocol i Run 14 APPENDIX A Use of the PiFOC The PiFOC is a focus correcting device It is useful for applications where drift in the z axis is anticipated E g super resolution imaging high resolution ima
9. el Frappa All Regions w frappa 466 11 If the Test FRAP doesn t work it will be necessary to modify the FRAP settings This is done in the Channels menu lees F Andor iQ 2 6 Andor Camera In the Left hand menu box select Channel Manager i from the Wizards wizards Process Analysis Plug Ins Help FH mai Select the Channel you want to modify i e a FRAPPA one And click Edit Channel Manager na Edit Copy Delete Close Channel Summaries Current Channel frappea_4ad 1 Channel frappa_4oo A Channel CSU 4o0 Quad e Channel CSU 488 5ingle F Channel CoL S6e Quad A Channel Col 405 Quad Click Next twice in the menu Pe Andor i 2 8 gt Andor Camne aa parr _ ap Sey You are now at the Bleach settings for FRAPPA menu File Image List Settings Wizards Process Analysis Plug Ins Help Setup Wizar Here you can control the Bleach settings Modify the dwell time i e how long the laser is on the sample and the Repeats how many times the laser visits the sample iol rappa iaa Setup FW and Shutters 3 Andor iQ 2 8 Andor Camera E loj xj File Image List Settings wizards Process Analysis Plug Ins Help lt lt Rack Next gt gt Cancel Setup Wizard Setup FW and Shutters for frappa 488 Control Settings Bleach settings e lt lt Back Next gt gt Ewell Time joo TE R
10. ging of structures in the centre of the cell long term imaging Do not have the cell you want to image in focus before you start this process as it may bleach Instead choose a region or cell which you aren t interested in acquiring data from and use this for calibrating the Z axis The PiFoc doesn t integrate into the Andor software it works alongside it To initialise the PiIFOC switch it on on the box Initialise 1Q2 first and wait until it is loaded Initialise the PiFOC software F I PI MikroMovet eR Agree when it asks whether to run the exe file The dialogue box below will appear Press Start to automatically connect to the controller iSPIMikroMove 2 9 0 4 Connections Tools wiew Help ILA kandeg Eala Fy E E fenter help search h i nect will connect Dares Lm Hast macros E 753 on RS5 232 COM One the PiMicromove the PiFOC controller has initialised the PiFOC will need to be auto Zeroed To do this choose E 753 Com 7 which is the PiFoc and select Start up axes PIMikroMove 2 9 1 4 0 x Connections E 753 COM Tools wiew Help a4 E Start Up axes ter EDE search her a g D Show Hide data recorder Show Hide wave generator Move multiple AHes os Host m macros C Piezo ch channels Cieno channels Configure trigger output Change command level ECA 80 000 75 846 gt Bt 0 100 80 000 on target 10000
11. have room for your experiment If not delete your old data 1 To switch on the microscope Note the objective needs to be in the focus feedback device older to be high enough to focus Switching on M I2 Ed Select the IQ2 icon on the computer to initialise 1Q2 Andor igQe Once the software is open e The stage may initialise e Put your sample on the microscope only when the Stage has finished initialising The software interface looks like this Here you control how Here you control the This menu is the one which The experiment is done Microscope itself Shows your image l Ea Camera DU8285_ P 5086 lolx 4 File c xy uias Analysis Plug Ins Help TCquisition Auntilia ry Devic m File Edit Imagelist Region MIP l e ay z ee pinNidt a i lt lt lt Tr E P lalwule i lem alala Riel golos Tije Live Snap CSU 568 Quad i r z saaie Acquisition Analysis RGB Analysis Movie Editor Spot Analysis i A eS Sas 2 Zheni ann Status Camera Selection Camera Jasi Sutter Yokogawa AOTF Trigger gt DU8285_VP 5086 Settings Exposure General Device Info Status E m Window Size 1004 1002 5 Gain J Full Chip Window EM Gain Enabled 297 RPTE een o m uto Save i PY Mi D User DatatAutolmagess E Shutter 7 gt Acquisition Calibration Open C Close Frame Transfer Live Fe 885_100Xoi 0 0666
12. he laser power e Select AOTF from the Acquisition Auxiliary Devices menu e Click on the laser you want to change Acquisition Auxiliary Devices e A pop out box will appear Adjust the laser power ALC Setings e Asa guide the lasers nm should be between 15 30 power Andor Laser Combiner Wavelength Laser Status e Ensure you hit Record at the end so the software remembers the am a settings Editing the protocol for channels and time Acquisition Lalipratia Ey Protocol Two channel Timelapse agn Image Two channel Timelapse 2 w Camera Binning Use Current 7 walt Allocation Off A Camera Selection Use Current H Events Fe Repeat T 60 times 5 sec i Move Channel CSU 499 Guad ioe 2 Snap Frame HAS Move Channel CSU 568 Quad 2 os Snap Frame Ease d End T 685_100xo01 006667 Edit repeat Repeats Interval sec T ce Once you have set up the acquisition channels and optimised the timelaspe protocol you are ready to Run the 3 Delta F experiment To do this press Run Aun Edit If you have made a mistake press STOP correct the error and re run the protocol AOTF T 2 E ioo l E iio Eh Wv il C FRAPPA t TIRF a There are general protocols which you can edit for your work You can also make your own protocol provided it is in your own user area i e not in the Ann or Timelapse setting which everyone uses this is so i
13. ion Use Current ah alle ES Protocol 2 Bun Se Channel CO bee es IC a el SES File Image list Settings Wizards Process Analysis Plug Ins Help CSU 568 Quad Acquisition Analysis AGB Analysis Movie Editor Spat Analysis Status Sas Auto Save To D User Data Mautolmages Lee Acquisition Calibratio 885_100x0i 0 06667 7 Protocol Two channel Timelapse agn Image Two channel Timelapse be a Camera Binning Use Current Ts Protocol F Run GP Channel Hesr Scan T Wal Allocation Off Camera Selection Use Current H S Events S d Repeat T 60 times 5 sec O E A Move Channel CSU 488 Quad Da Snap Frame H A Move Channel CSU 568 Ouad LT Snap Frame ae d End T Pq Device Setup Loop Protocol W Display Image M Show All ila Taking an movie The spinning disk is a hybrid between a wide field epi fluorescent microscope and a point scanning confocal The disk itself makes the points of confocal light so the scan speed doesn t need to be controlled Acquisition is controlled very much in the same way as in Metamorph so gain and exposure are the same It is also possible to frame average and to change the laser power to improve your image Send the light to the camera Knob on right hand side of microscope turn towards you until it click into place turn back slightly so the light is centred in the camera Once you have seen y
14. mewhere ag Camera Binning Use Qurrent random T Wait Allocation Of Fa Camera Selection Usel lurent F Run AP Scan test scan Flee Events SE Repeat T 61 times 20 sec G E eed Be Move Mert eee Guad fg Snap __ Delete T Loop Protocol i Display Image The XY scan map will appear XY Scan Map test scar lol xi 77 050 77 100 77 150 C Fields C Current Position Current Position Rescale Copy to Clipboard IL seve Legend 5 gt Andor iQ 2 8 Andor Camera 4 lol x Fie Image List Settings Wizards Process Analysis Plug Ins Help Setup Wizard Setup XYZ options It is possible now to either select a pre recorded scan and use these xy settings or edit them Please don t edit the 12 and 24 well plate presets Select Z Device Device Setup Next gt gt Multi_Field options To edit the positions use the following dialogue Select Number of Fields E 1 Select the number of positions you want Press NEXT FIELD to select position and move to next field Press PREVIOUS FIELD to select position and move to previous field 2 Move the stage using the joystick to the x and y position you want this is recorded in the x y and z position window Please ensure image is focussed as z position will be recorded if available x position 77091 osition 3471 6 a Current Field 1 z position 49 99 lt lt Previous Fiel
15. nnection MICRONS Z stacks can be added into a protocol by adding a time repeat into the Right click and edit More channels can be added by right clicking below the Frame option and adding a channel Ensure that the End Z item is after the new channel otherwise the software will do a z stack of the first channel and either not one of the second Or alternatively a z stack of channel 1 and then a z stack of channel 2 The problem then is the two channels may not correctly register Choose Run to collect the data a Run SUS ered E D MUser Data Aubol mages a Acquisition Calibration 7 Protocal 2 stack Ag Image stack pe 2 Camera Binning Use Current Wait Allocation Off m Camera Selection Use Current fee Events lf Repeat 2 3 6 um in 44 planes oo Qe Move Channel CSU 568 Ouad Alsou fai Snap Frame w Centre 2 Scan 365 100X o1 0 06667 Protacal Ta Protocal G Rur Z Channel Ee i Peds Insert Delete op Frotocal M Display Image I Show All NB Please come and ask if you need a new protocol setting up as the BALM manager is always happy to support these type of requests 10 To do a FRAP experiment Select a protocol with FRAP in it If you haven t done FRAP for a while you will need to Protoca do a test FRAP to ensure the laser power is still OK for your experiment a H Protocol To consider when carrying out a FRAP experiment
16. ontrol window for the PiFoc will appear s E i Axis 1 2 P 725 1CD c State on target Set the target in this window to 80um The Pifoc can move between 0 and 100um but 80um means thatthe a device can sense the z axis position accuractely but still is within its ie range of movement to correct any drift upwards or downwards a ref mf Refocus on your sample on the microscope um Target fo 000 4l gt Find the cell you want to collect data from and set up the experiment fa po pomo 16
17. our specimen you change the dropdown menu to select your laser for instance CSU 488 Quad Make sure to set the light so it s going to the camera on the microscope Hit Live to visualise the image on the computer screen To save your settings for each channel click Record Each time you change any of the acquisition parameters you need to record it or the machine won t remember Controlling the camera Change the EM gain no more than 300 DU8285 YP 5086 Settings Exposure General Device Into Status a8 Gain W EM Gain Enabled 297 4 Exposure Time Frame Transter Live af Readout mst 32 723 Frame Fiat Actual Hate 0 00 For a nice picture use frame averaging the amount of averaging needed varies between samples so you need to optimise it yourself Ensure the Use box is checked hutke _ 6 Open f Close Frame Averaging Change Exposure time f Display Every Frame NB for live imaging ensure that the time needed to acquire and Display 1 Frames s image which is the exposure time X the number of frames averaged is less than the rate of data acquisition Trigger Contra Ensure you hit Record at the end so the software remembers Intemmal Trigger Ext Trigger Timeout ras 1000 External Start PC RAM MB 2047 the settings Camera Staty Camera is working Tips a Temperature 7U To change t
18. rolling the image histograms appears Left clicking allows you to adjust the offset black point and right clicking allows adjustment of the maximum intensity displayed This doesn t alter the rs raw data in your image just what you see It can be Bee helpful to adjust this for dimmer samples gt Image t ast Options View 2j E 5388 000 Max E 280 0000 Min k_ 000000 Step Gamma Coefficient 7f Automap Off F Smooth M Use Region of Interest i Cancel F 99 1 Contrast i Wap on mouse up a SAVE YOUR DATA To save data click on the file menu top left in the image itself and gt Camera 1 DU8285_ P 5086 choose save as tif File Edit Image list Region View Fast MIP Looking at your movie after acquisition The Wavelength dialogue allows you to switch off the display of channels 4 d j Time Wavelen th The Time menu allows you to pla EGE ESSE TST m premen a em through the movie i CE B fo EE a aliaj m pi ee r aa EN oe Flatten Time ez s Automen 100 23586 Image List Once you have finished collecting data go to the Image List o gt Camera 1 DU8285_ P 5086 Image List Region View Fast MIP ier alsalaf Nel The Image list menu will appear P32 Image Manager D KineticImageDisk D7WKRN4J Administrator File Edit Comments Options Image Details Name Time
19. t a y Repeat lt AB um in 44 Lica O ES Mowe Chanel LELLES Oued a 7 7 entre can If Centre Z scan is selected the centre will be assumed to QE i Snap mee o B z be the plane in focus Delete op Protocol M Display Image I Show All 3 In the protocol select and edit the Repeat Z File Image List soar PERE Process Analysis Plug Ins Help amore j Setup Wizard i This makes the Z scan menu appear Setup xyz options Select Z Device The top and bottom of the Z stack should be chosen by moving the Device Setup piezo focus drive using the dialogue at the bottom of the menu Next gt gt Cancel Z Series options SetStat 4569 SI Set Centre 50 09 Zi SetEnd 54 49 X adjusted AZ um 0 2 Nyquist Cubic Planes 5 ack Definition AZ Number of Planes C Z Positions Having navigated to the top start of the cell using this m pick Set Start Then navigate to the bottom of the cell using the piezo control menu and pick Set End Select the space between Z slices by using dZ To obtain optimal lee E resolution select Nyquist double over sampling Alternatively the number of planes can be chosen If there is a problem with the piezo Z drive and it needs resetting picking the Piezo Menu option Refocus to where you need manually and reset in the menu Acguisition Auxiliary Devices kep Settings Co
20. t isn t confusing for less experienced users 1 Set the Interval of the time lapse i e frame rate This is done by right clicking the interval icon and editing the interval length in the pop out box You put the number of repeats and interval between images in the edit repeat box 2 If you wish to change a channel right click in the box and select change Channel you can also insert and delete elements in the protocol here te Snap Frame He Move Channe Move To n a Snap Frame Adjust Camera Acquire Insert Delete Change Channel Changing the brightness and contrast on the camera during r imaging AIOC EE Ski m o s T d x ro H With Fluorescent proteins which are easily bleached it can be better to alter the part of the image histogram shown on the camera while imaging This way the laser power and exposure time can be kept low But you can still see and focus on your image It works like autoscale in Metamorph To optimise the Brightness and contrast press manual control which is at the bottom of the image window aLL L e z Wavelength T him s ms Frame Number Minrange Maxrange Increment Interval sms a Min range Masrange Increment K ae FR fF ono S fo 24 fi EEE Tila _Setbin Serma BK I Automap 1000 754101 858 1982 1113 x e w 00 2 Set Min Set Max Frame Humber E oe Flatten Time WM Autormap A dialogue box cont
21. use is hovering over ifdicated by a white arrow on the image This can be useful when setting the lawer power and gain and exposure of the camera If you want to use the focus feedback controller PiFoc see Appendix A at the back of the manual 2 To look at your samples J microscope away from you the right hand side of the microscope stand Camera Status Wait 5 minutes for the camera to cool to 70 C Camera is working Temperature 70 Using the Acquisition Calibration box select the objective to be used for this imaging session andor i 6 Andor Lamera E LLa Fie Image List Settings Wizards Process Analysis Plug Ins Help The system won t work if this doesn t happen me Ei 38 ei P Open Imaris Save Live Snap CSU 568 Quad Acquisition Analysis AGB Analysis Movie Editor Spot Analysis Status ee Auto Sav a DAU ser Data Autolmages E Acquisition Calibratio FH 885_100Xoi 0 06667 2 a a E m N Ensure the light is going to the eyes turn the knob on the RHS of the Bright field can be turn on by pressing the switch on Open the fluorescent shutter on the Mercury halide box Select GFP Eyes RFP Eyes or DAPI Eyes from the Acquisition Auxillary Devices menu the middle menu a E i A See T En Tran l hal 0 op Camera Selection Sutter Yokogawa AOTE Trigger 4 gt DUS285 VP 508
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