LED intensity

[Xinchi]

Hi Daniel,

We have successfully built up our system! However we have one question regarding of the LED system.
We found out when we first connect our microscope to the mouse, we could observe good quality of neurons (clear white dots). But after a while (maybe 30mins), the image starts to become blurry. We used the intensity of 20% with GCamP6m. We tried to lower the value but the same problem happens. I am not sure this was because of GFP bleach.
Could you share some tips or do you have any good recommendation value of the LED intensity for GRIN Relay Lens system?

Many thanks!!

Xinchi

[hoogland]

Hi Xinchi,

An intensity setting of 20% seems way too much light. We typically use 2-3% to get decent recordings.

Cheers,

Tycho

[Daniel Aharoni]

30 minutes of 20% excitation will very likely result in significant photobleaching. Consider increasing the GAIN setting in the Miniscope DAQ software to allow for lower excitation power.

[Xinchi]

Hi all-

Thank you all for the advice! We will try lower the intensity then.

Bests,

Xinchi