GCaMP virus

[Ana Carolina Bottura de Barros]

Hi everyone,

A few questions regarding viral injections of GCaMP:

Has anyone tried imaging GCaMP7 injected mice? How did that go?

Also, regarding GCaMP6, is there specific one (s, m, f) that seems to work better?

To those, based in Europe/UK, where do you order your virus from?

Thanks!

[Richardson Leão]

gcamp7 is the brightest but they are less available in vector cores (https://www.nature.com/articles/s41592-019-0435-6). the s,m,f will depend on your application. for spikes, the gcamp6f is already slow but it has the fastest response, so we only use gcamp6f. charite berlin has a vector core that has gcamp constructs. camk2-gcamp we buy from addgene (at 3 times the price)

--
You received this message because you are subscribed to the Google Groups "Miniscope" group.
To unsubscribe from this group and stop receiving emails from it, send an email to miniscope+...@googlegroups.com.
To view this discussion on the web visit https://groups.google.com/d/msgid/miniscope/d4754177-c303-40be-8cc5-97a29de2615dn%40googlegroups.com.

--

Richardson N Leao, MD PhD,

Neurodynamics Lab, Head

Brain Institute, UFRN/Brazil

[cke...@gmail.com]

We have found that for CA1, we get the most reliable and strong expression using AAV9.

[Matthias Klumpp]

Am Mo., 12. Okt. 2020 um 11:51 Uhr schrieb Ana Carolina Bottura de
Barros <anabo...@gmail.com>:

I am using GCaMP-7f AAV1 from AddGene for CA1, and it works very well
for both injections and coated lenses.
(But it looks like AAV9 would be worth trying as well).

--
I welcome VSRE emails. See http://vsre.info/

[Mahesh Miikael Karnani]

Hi Ana! I've used GCaMP7f with synapsin promoter from addgene. It's fantastic!

[cke...@gmail.com]

I should clarify that we use the CaMKII:6f virus that's marked as "UPenn" on addgene. Presumably Synapsin gives interneuron expression also?

[Mahesh Miikael Karnani]

Yes it does!

[Zeeshan Haqqee]

We use gcamp6f with either CaMKii or h-syn promotor as well, and they both seem to work great for CA1. 

On a related note, for how long and at what LED intensity do most people record with (either on v3 or v4?). I've been trying to figure out how long I can safely do in vivo recordings in CA1 with gcamp6f without significant photobleaching. Have tried as long as 30-40 minutes per day at LED 10-18 (on the v3 so far) and I *think* the photobleaching effects are minimal, but it's hard to tell (esp. for hippocampal cells). 

Zeeshan

[Ana Carolina Bottura de Barros]

Hi everyone! Just wondering from people who use GCaMP7, what dilution do you use? Does it change from s, m and f?

Thanks in advance!

[Md. Mamun Al-Amin]

I purchased AAV9.GCaMP7f with synapsin promoter (2x10e13). I have not tested yet. I would like to know what concentration it will work?

Has anyone tried with coated lens?

Thanks

Mamun

[Zachary Zeidler]

I've compared 7f undiluted (titer of 2e13) to a 1:1 dilution, and the 1:1 dilution seems to be better. Less overexpression and less permanently bright patches, but still a robust signal. 

And a side note related to GCaMP -- Addgene now has vectors for the GCaMP8 line (serotypes 1/2, synapsin promoter, both standard and retro). GCaMP8f has 4x faster rise and 2x faster decay than 7f, and 8m has 3x the sensitivity of 7f with 4x faster tise. It's easy to fall into the trap of always chasing the next best tool instead of actually doing experiments, but if you're about to start out a new project, it might be worth checking out the GGCaMP8 line. Haven't tested them out but I might soon :)

--

You received this message because you are subscribed to the Google Groups "Miniscope" group.
To unsubscribe from this group and stop receiving emails from it, send an email to miniscope+...@googlegroups.com.

To view this discussion on the web visit https://groups.google.com/d/msgid/miniscope/7aafc122-766d-4f99-9dec-81ef07c72af6n%40googlegroups.com.

[Jyun-you Liou]

I have been using GCaMP7f at the cortex. It works! 

[Ana Carolina Bottura de Barros]

What dilution do you use?

Thanks!

You received this message because you are subscribed to a topic in the Google Groups "Miniscope" group.
To unsubscribe from this topic, visit https://groups.google.com/d/topic/miniscope/N4p9b_i51og/unsubscribe.
To unsubscribe from this group and all its topics, send an email to miniscope+...@googlegroups.com.
To view this discussion on the web visit https://groups.google.com/d/msgid/miniscope/ca5c2456-6889-4c51-8b50-b523c07535d4n%40googlegroups.com.

[Jyun-you Liou]

I used 4 x 10^12 titer. The injection method was the same as the NINScope paper.  Up to now, they all work from 1 x 10^12 to 4 x 10^12.  I found the key is to ensure good contact between the glass and the brain to avoid scattering and uneven surfaces.