Using Miniscope V4 to image somatosensory cortex in mice

[Jyun-you Liou]

Hello folks,

I am a very new Miniscope user. I just received my first Miniscope v4 from OpenEphys a few weeks ago. My project is about imaging the somatosensory cortex. The animal I have is Thy1-GCaMP6f mice. 

Our lab did the surgery and put a 3 mm cover slip over the cortex. We were hoping to see some cells. However, although we can see very clear capillary, even things look like red blood cells moving in the vessels, we can not see individual neurons. All we saw was a very right background fluorescence, fluctuating according to the animal's behavior/some sensory stimulation. I also tried to move my focal plane.  I saw very clear brain surface vessels at about +60 EWL depth.  I scanned over all depth down to -127.  I still couldn't find isolated neurons.  They were all bright, blurry, and dynamic fluorescence.  

I do not have a specific layers in my mind to target at, as long as the cells I see are excitatory cells. I have read some discussion threads here. It looks to me suctioning the cortex is not a good idea. I have also done some literature review.  Considering the geometry of the somatosensory cortex, it looks to me people some people have been using microprisms.  

Can please help me figure out what I should do to get single cell fluorescence in this situation? I am really new in this field and my lab is not an expert in cellular imaging.  Really hope that I can get help here. 

Appreciate any suggestion. 

[Stephen Larson]

Hi there,

    Did you manage to solve your imaging challenges?  I know of some other folks who are trying the same thing you are.

Thanks,

  Stephen

[xinman yin]

Hi，stephen

    I am also interested in this direction. Would you like to share the cutting-edge knowledge of this direction with me or exchange some knowledge with me? I am a first-year graduate student and a novice in this field.

[Yukti Vyas]

Hi all, 
Have you tried checking with viral injection or viral-coating cranial windows in the somatosensory cortex? And if so, are you getting the same issue?  I wonder if modulating the concentration of the injection and level of expression might help get single cell resolution rather than a generalised background flurorescence.  Please let me know if you have tried this :) 

Best,

Yukti Vyas