Unsuccessful imaging through cranial window in Thy1GCaMP mice with V4

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Amit Koren

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Nov 16, 2022, 4:36:55 AM11/16/22
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Hello,

Despite many controls such as double-checking the same mice in the 2-photon and imaging constant expression of microglia in CX3CR1 mice, I’m having issues imaging neuronal activity through cranial windows in Thy1GCaMP6s mice with the V4 Miniscopes.

I tried many variations of cranial windows, coverglass depths, and cortical areas. 

My goal is to image in the least invasive way possible, so I would be rather not implant a relay lens. 

I suspect that I am still not reaching a sufficient depth to image L2/L3. Has someone resolved a similar issue?

Thank you for any advice/suggestions! 


Isaac Anthony

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Nov 17, 2022, 12:36:48 AM11/17/22
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Hello,

I have not heard of anyone successfully resolving cells in the cortex of transgenic GCaMP mice (myself included after much effort). Deeper regions like CA1 work with transgenics if I remember correctly. I switched to AAV injections of GCaMP6f and successfully imaged cells in somatosensory cortex. Happy to share more if no one else in the group has had success with imaging cortex in transgenic GCaMP mice.

Amit Koren

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Jan 18, 2023, 5:18:55 AMJan 18
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Dear Isaac,

Thank you again for your helpful reply, we took your advice and switched to viral injections.

I wanted to know if you have any tips regarding successful cortical imaging through cranial windows? And/or if you could share a general outline of your injection and surgery protocols?

Thank you very much,

Amit
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