WangLab:Confocal Microscopy Usage

Confocal Microscopy Processes:

Note

• Do not touch the fiber cable for Laser!
• If you want to change objectives, see Jerry!

1. Turn on Arc Lamp first.
2. Turn on Laser Power, Make sure the laser is on Standby mode (Standby light should be on). Turn the key to On->Start->On on laser.
3. Turn the key on the laser set besides microscopy to On (Off->Spinning->On).
4. Turn on computer, Turn on camera, turn on controllers for Stage, filter shutter
5. Get into Simple PCI, Select file->manage profiles->Simple PCI 5.0
6. Click on the camera icon, Choose Monochrome for single staining (you can choose other options based on your exp.), Select filter set, for FITC, it is 488/525.
7. Only one small drop of oil on lens (must be Olympus oil), mount sample slide, Turn on the regular light lamp, focus the sample with eyepiece.
8. Turn off regular light, switch to SP (side port?).
9. Turn the switch from Standby to On on the shelf laser controller.
10. After experiment, turn the key on the shelf to Off, the key besides the microscopy to Off (On->Spinning->Off).
11. Lower Objective, take off sample, clean lens (wipe off the oil excess with dry lens paper, and clean the lens with lens paper with lens cleaning solution).
12. If the noise from laser is gone, turn off the power switch of laser on the shelf.
13. Turn off light lamp.
14. Change simple PCI profile to Null
15. Turn off camera, controllers for shutter and stage.
16. Turn off computer after image analysis.