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Troubleshoot: I cannot align the laser

If you cannot align the laser it can have several reasons

The laser spot is not or hardly visible

  1. Take out the IR filter by removing the DIC slide of your microscope
  2. Change to a magnification of 10x or higher
  3. Adjust the exposure time in the settings dialog
  4. Check if there is an air-water interface between probe and probeholder. Add more liquid medium if so.
  5. Check if there is a bubble on the probeholder. Remove it with a kimwipe if so.
  6. Check if there is condensation fog on the probeholder. Use soap water or an anti-fogging agent on the inside of the probeholder and dry it properly

 

The laser is perfectly centered on the probe but the signal quality is still "poor"

You should always get a signal with FluidFM probe. Probably the AFM was used with regular AFM probes before, which have a different geometry. The photodiode needs to be adjusted now.

  1. Adjust the photodiode with screws Bx and By (the two screws to the left) until you get a good signal quality.
  2. If the prior step does not help. Adjust the position of the laser a bit further away from the probe opening ("up" in the microscope image). Try step 1 again. 
  3. Check if the CytoClip is properly mounted on the probeholder. Try step 1 again.

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