Spinning-disk X1 troubleshooting
I want to locate my sample though the ocular but there is no fluorescent light coming out of the objective
- Check the lamp is switch on, the optical configuration you selected (“DAPI”, “GFP-B”, “TxRED”) and you click on the button to switch on the shutter
- Check the FRAP mirror, at the back is well in its working position (the black rod on the red little box at the back of the microscope should be at 45°)
- Check the diode at the front of the fluorescent lamp
- If it is blinking, it means it didn’t initialize properly. Restart the fluorescent lamp
- Check the white Nikon remote next to the keyboard, the “ND” diode should be set on 1.
- Check the camera lever on the right side of the microscope, it should be oriented toward the desired camera
- Check the Metamorph version you used, as it should match the desired camera selected previously
- Check the sample focus through the ocular before coming back to Metamorph
- Check the Z-speed near the joystick : it should indicate “Coarse”
- If no movements are visible, switch off Metamorph, and reboot the multisocket below the keyboard. Restart Metamorph.
- Remove and clean the slide
- Clean the objective
- Add immersion medium on the objective (if oil objective)
- Put it back on focus
- Add a folded Kimtech tissue between the stage and the sample holder to tight it well.
- The Kimtech tissue raises a side of the sample holder, or the sample holder is not firmly set at its lowest point. Readjust it to minimize this effect.
- Perform a Köhler alignment
- Do not use the well on the edge of the multiwell plate, as the objective can’t reach them