Forum - Experiment length using gcamp6f

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[#41]

Hi Everybody,

When using AAV1.Syn.GCaMP6f.WPRE.SV40 from Penn Vector how long are people able to record before the signal bleaches out? How long do you have to wait for it to return?

I found [1] but it is for 2 & 3 photon imaging so I'm not sure how relevant it is.

Best, Alex

Posted by Alexdunlap on 22 April 2016 at 15:33.

Hi Alex! From our experience, when we record CA1 for 10-15min per day, every other day, we see little to no photo bleaching. With our miniscope, the LED intensity is typically very low, about 2-5% on average. We start to see bleaching after 30 minutes and it seems to recover after a day or two. The trick for using such low LED intensity is turning up gain. This seems to work well in areas where the cell population is dense, such as CA1 but may be different in other brain regions. Let us know what your experience is too! Good luck!

Posted by Denisecai (administrator) on 22 April 2016 at 22:52.

Thanks! I'll let you know what we get as well.

Posted by Alexdunlap on 25 April 2016 at 17:19.
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