My lab ordered GCaMP 8f a while back, but during imaging there was not a lot of activity (1:10 dilution in PBS) in the ~10 mice we tested. We cannot use 8s because we are trying to image PV neurons, which have very fast spiking activity so we want as much temporal resolution as possible. Has anyone used GCaMP8f successfully? We were just wondering if we got a bad batch somehow or there was a different dilution we could try.
Hi Christian,
What is the titer of the batch that you ordered? Maybe with a 1:10 dilution its to low? How long did you wait after injecting the virus? Where did you order the virus and what is your sampling rate?
Hi Christian,
In our hands Gcamp8m works very well at 7.7 x 10E12 vg/ml for fiber photometry. See if your concentration was maybe too low. Also what excitation wavelength do you use? Also just in case at the 2P you should use 900-920nm and not 800 for Gcamp.
Hi Christian,
Have you had any luck with your virus at a different dilution?
We are having a similar problem with the cre-dependent GCaMP8s we ordered from addgene at the end of 2021. We are imaging pyramidal neurons using 2P through a GRIN lens. We have not diluted and I believe the titer is around 2.9 x10E13vg/ml so we initially were thinking that we have too high a concentration. I am about to try a lower concentration. I have a colleague who has said that FLEX GCaMP8f worked well for him at 1:10 dilution, although has not tested it extensively. I am also worried about a problem with the virus batch so it would be great to hear if you have had any success!
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