Imaging of interneurons

Florian S

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Dec 8, 2021, 7:00:22 AM12/8/21

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Hello everyone!

We are currently trying to image interneurons using the Miniscope V4.

Does anyone have experience with this, specifically regarding PV cells?

I would be very happy about any shared experience!

Best regards,

Florian

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Florian Steenbergen
Albert-Ludwigs-Universität
IMBIT - Optophysiologie
Georges-Köhler-Allee 201
79110 Freiburg
Germany

xiaoyu peng

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Dec 8, 2021, 8:04:42 AM12/8/21

to Florian S, Miniscope

I tried VIP and SST in cortex, it looks great with V4! I didn't prepare PV neurons in the current cohort, but I was able to see some with V3, I can only expect better with V4.

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Florian S

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Dec 8, 2021, 8:19:11 AM12/8/21

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Hey,

thanks so much for the quick response! Would you mind sharing which version of GCaMP you used? I currently aim to use GCaMP8f or 8m, do you have some experience with these?

And is there a possibility that you share a brief video example so I can get an idea of what signal I can expect for each interneuron type?

By advance, thank you very much and best,

Florian

GERMANY

xiaoyu peng

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Dec 8, 2021, 7:31:15 PM12/8/21

to Florian S, Miniscope

I used AAV1-flex-GCaMP6f, I haven't tried 8f, could you share the experience when you try it, would love to know. I attached SST neurons video, I only have these at hand. they have these large domain lightening up that I didn't see in other neuron types, in addition to cell body signal. I enhanced the brightness to visualize better, the real video was dimmer, but because the noise is pretty low, the analysis works pretty well. --Xiaoyu

15 -SST.avi

Philip Vassilev

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Dec 8, 2021, 8:24:53 PM12/8/21

to xiaoyu peng, Florian S, Miniscope

Hi,

Thanks a lot for sharing this video! I have seen this intense fluorescence over a large area as well, and I was wondering if it was an artefact or actual activity. I am, however, using a synapsin-driven vector, so I don’t know if it’s coming from interneurons specifically. Do you have a reason to expect these large increases in fluorescence from interneurons?

Kind regards,

Phil

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Richardson Leão

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Dec 9, 2021, 4:22:08 AM12/9/21

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has anyone used mDLx-GCamp for interneurons?

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Richardson N Leao, MD PhD,

Neurodynamics Lab, Head

Brain Institute, UFRN/Brazil

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Florian S

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Dec 14, 2021, 1:47:32 PM12/14/21

to xiaoyu peng, Miniscope

Thanks so much for the example, it is very cool to gain some better idea of what to expect.

Best,

Florian

Am Mo., 13. Dez. 2021 um 14:23 Uhr schrieb xiaoyu peng <xiao...@gmail.com>:

Here it is, imaged by V3. I downsampled and enhanced brightness in image J. PV neurons seem to be " slow", I remember I checked the signal after baseplating and was thinking why some cells are bright for so long, are they dead? I was used to watch excitatory neurons. The first time I watched this PV neuron mouse, I had a moment thinking something was wrong with it. It all makes sense once I think a bit more about their spiking features. --Xiaoyu

On Sat, Dec 11, 2021 at 2:35 AM 'Florian S' via Miniscope <mini...@googlegroups.com> wrote:
Thanks everyone for the input and thanks for the video @Xiaoyu, looks awesome! I am happy to share experience as soon as we gathered some.
If you happen to get your hands on a video example of the PV cells from the V3 recordings I would be super grateful!
If anyone else has experience with imaging of PV or SST cells, I would be very interested in sharing knowledge on this.

Best regards,
Florian

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xiaoyu peng

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Dec 14, 2021, 1:47:33 PM12/14/21

to Florian S, Miniscope

Here it is, imaged by V3. I downsampled and enhanced brightness in image J. PV neurons seem to be " slow", I remember I checked the signal after baseplating and was thinking why some cells are bright for so long, are they dead? I was used to watch excitatory neurons. The first time I watched this PV neuron mouse, I had a moment thinking something was wrong with it. It all makes sense once I think a bit more about their spiking features. --Xiaoyu

On Sat, Dec 11, 2021 at 2:35 AM 'Florian S' via Miniscope <mini...@googlegroups.com> wrote:

Thanks everyone for the input and thanks for the video @Xiaoyu, looks awesome! I am happy to share experience as soon as we gathered some.
If you happen to get your hands on a video example of the PV cells from the V3 recordings I would be super grateful!
If anyone else has experience with imaging of PV or SST cells, I would be very interested in sharing knowledge on this.

Best regards,
Florian

richard...@neuro.ufrn.br schrieb am Donnerstag, 9. Dezember 2021 um 10:22:08 UTC+1:

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msCam14_PV_small.avi

Jessica Froula

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Sep 13, 2022, 11:53:42 AM9/13/22

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Hi Florian,

Were you able to image successfully using Gcamp8m or 8f in interneurons? I am gearing up for some new experiments and thinking about switching from 6f, and I would be quite curious to know how it went for you.