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Dear all,
We have a user working on mouse brain. He is trying to image calcium transients from mice injected with GCaMP6m and implanted with a GRIN less in a deep brain area.
The microscope is an inscopix miniature microscope mounted on the skull of the animal and the GRadient INdex (GRIN) lens is the following: 500 um diameter, 0.5 NA, pitch 2.
He consistently observed that the best focus with this lens is 500um above the top surface of the GRIN lens, but unfortunately this is not where the most of the cells are.
If he goes closer to the GRIN lens and focus 300um above the top surface, he can see activity from many more cells but they are out of focus.
So the placement of the implanted GRIN lens is not optimal in relation to the area of the brain where there are more infected cells.
How to determine if the plane where we have the maximum number of cells right now is too close or too far away from the bottom surface of the implanted GRIN lens? My guess will be that he needs to make sure that the GRIN lens is 500um from the plane he wants to observe - meaning deeper. But this is a lot of work for him to test, so we would like to double-check before he plans the next surgeries: Should he should implant either higher or deeper in the brain?
Does anyone had a similar experience with GRIN lenses and this miniature microscope?
Many thanks for your help,
Best regards,
Alexia
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Dr Alexia Loynton-Ferrand | Research Associate in Advanced Light Microscopy | Imaging Core Facility | Biozentrum, University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
Phone: +41 61 207 57 30 |
[hidden email]<mailto:
[hidden email]> | www.biozentrum.unibas.ch<
http://www.biozentrum.unibas.ch/> | www.microscopynetwork.unibas.ch<
http://www.microscopynetwork.unibas.ch/>
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