Transcript
Raman confocal imaging microscope
Instructions for Horiba Raman confocal imaging microscope
Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Dianwen Zhang (Email:
[email protected])
Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
Revision History
Authors and Editors Dianwen Zhang Dianwen Zhang
Revision Date 4/20/2012 12/22/2014
405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
Table of Contents The Raman confocal microscope system: Overview ................................................................... 1 Laser Safety Requirements .................................................................................................................. 2 Start up the microscope system ......................................................................................................... 3 System configuration for general users .......................................................................................... 6 Front panel ............................................................................................................................................................. 6 Labspec software ................................................................................................................................................. 6 Shut down the system ............................................................................................................................ 7 Troubleshooting ...................................................................................................................................... 8
405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
The Raman confocal microscope system: Overview The left rear view 830 nm laser
785 nm laser
405 nm laser 633 nm laser
532 nm laser
The front view
Andor Newton EMCCD camera
Synapse camera Fine focusing Coarse focusing
Laser button Objective turret
Halogen lamp for epi‐illumination
405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
Laser Safety Requirements The Raman confocal is a Horiba LABRAM product and a class 3B laser product, which means Laser beams and reflected beams could be dangerous. Diffused reflected beams in most cases do not present any problem. Lasers are high intensity light sources producing visible or invisible light (for example, 785 nm and 830 nm lasers in the system) at specific wavelengths. This concentrated energy in a narrow laser beam may cause damage to biological tissues, especially to eyes. All below labels on the microscope indicate a hazardous situation where there is a risk of serious injury to be caused by laser radiation. Special attention must be paid.
Before removing or changing objectives, the laser button on the microscope front panel must be in “off” position. In case of removing objectives, the empty objective positions on the objective turret must be covered with objective caps. When using 785 nm and 830 nm lasers (invisible), all users are required to wear laser goggles throughout experiments. 405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
Start up the microscope system 1. Turn on the laser. The laser controllers are shown in Figure 1. Only one laser is supposed to be used at a time; Do NOT adjust the settings on the laser controllers. For the 405 nm laser (Figure 1(a)), plug the power supply to the nearby power outlet. For the 532 nm laser (Figure 1(b)), turn on the key switch, and press the “Enable” button when it starts flashing. For the 633 nm laser (Figure 1(b)), turn on the key switch on the laser controller. For the 785 nm laser (Figure 1(c)), turn on the key switch located at the rear of the laser. For the 830 nm laser (Figure 1(b)), press the power button on, wait for 30 seconds and press the shutter “on/off” button on. (b)
633 nm laser controller
532 nm laser controller
(a)
Power
830 nm laser controller
Shutter
785 nm laser “on/off” switch at the rear of the laser. (c)
Figure 1 (a) 405 nm laser power supply; (b) 532, 633 and 830 nm laser controllers; (c) 785 laser
2. Setup the laser path to microscope Following the note on the yellow label beside each plunger (#1‐4 in Figure 2), set up the four front plungers to guide the laser to the laser entrance port of the microscope. 405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
Laser entrance (a hole) to the microscope Plunger 2
Plunger 1
Plunger 3
Plunger 4
Figure 2 The front plungers
For the users who use 633 nm laser and don’t use Superhead, please also check the fifth plunger (Figure 3) in the system:
Plunger 5 needs to be pulled up for the use of 633 nm laser when using the microscope.
Figure 3 The rear plunger
3. Check the two optical filters inside the sliding chamber (Figure 4) on top of the microscope. If the labels of the installed filters doesn’t match the laser wavelength, change them.
Open the chamber carefully
Figure 4 Filters chamber
405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
Carefully change these two filters if necessary.
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
4. Turn on the LED lamp for epi‐illumination.
Figure 5 LED light for epi‐illumination white light
5. Open the control software – Labspec and load sample for measurements. If Labspec freezes or gives an error message “Connection with S3000 fails,” follow the instruction at page 8 to restart the microscope system.
405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
System configuration for general users Front panel Push in the
Push in for microscope mode
DualScan/Standard plunger
Pull out – only Synapse for general users
Use microscope mode
Turn it on only when laser is needed
Use only Synapse for general users
Optional
Optional
Use only XY stage for general users
Labspec software For the general use of the Labspec control software, please refer to Chapter 9 of the software manual (http://itg.beckman.illinois.edu/microscopy_suite/equipment/raman_imaging_system/Lab Spec%205_manual_for_general_users.pdf).
405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
Shut down the system 1. 2. 3. 4. 5.
Close the Labspec software, and log off the computer; Turn off the LED lamp (The power button at the rear of the lamp); Turn the “Laser” button on the microscope front panel to OFF position. Turn off the laser that you have used; Clean up the working place (Take away all your stuffs; used glass slides or coverslips should be thrown into sharps waste bin; etc.)
405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
Troubleshooting 1. Problem: When starting Labspec, it becomes frozen or shows the below error message before the final main window can appear.
Solution: Follow the below steps to restart and reset the microscope. 1) Close Labspec. Sometimes you also need check the Windows Task Manager to make sure that there is no a process called “NGSLabspec.exe” running in background. 2) Restart the microscope. Find the below socket and the microscope power box beside the laser controllers on the optical table. Turn off the socket switch pointed by the yellow arrow. Wait more than 1 minute and then turn it back on. Now the microscope is restarted.
3) Start Labspec. Labspec should run normally, and after a few common device initialization, it will show the below stage calibration message. Before you click “OK”, make sure that no sample is loaded or the loaded sample won’t hit the objectives when the stage moving in its full range. Click “OK” to start the stage calibration.
4) Initialize the three key microscope components ‐ “Hole,” “Slit” and “Spectrometer”. See the below image. Click the red thunder button at the top‐ right on the “Hole” control panel, wait until the initialization process is complete, and then separately repeat this procedure for “Slit” and “Spectrometer”. 405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana‐Champaign
Operation manual of Raman confocal
2. Problem: Labspec can start without errors, but will malfunction. Solution: Follow the below steps to reset the software. 1) Find the configurator icon on the toolbar of Labspec. Click on it to open the below configurator window:
2) On the configurator window, click the button “Load”, and then find and load the default setting file “Default.ngp” under the folder “Manual” on your desktop.
405 North Mathews, Urbana IL 61801, USA Phone: 217‐244‐0170; FAX: 217‐244‐ 6219
http://itg.beckman.illinois.edu/
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