For example, you inject AAV-GFP particles into the mouse cortex, and get excellent GFP fluorescent signal a few weeks later.
But can you detect the presence of GFP mRNA or other AAV related transcripts in single cell by qPCR or other RNA sequencing techniques?
Yes, but if you are interested in identifying which cell(s) are expressing the mRNA, you will have to do an in situ hybridization or something analogous to it.
Hi Jiahao,
without being an expert in the field,
You have to manage to isolate the single cell first, then you'll be able to measure GFP.
The best way to isolate single GFP+ neuron so far is after electrophysiology recordings, from a slice prepared for that
http://journal.frontiersin.org/article/10.3389/fgene.2012.00124/full
However, this is very challenging technically
Maybe laser microdissection might be possible, but I'm not sure.
Hi Jiahao,
You can dissociate your cells and do a single cell RT-PCR on individual cells. You may also you brain slices but personally I would recommend dissociated cells. The idea is that you will use a micropipette controlled by micromanipulators under visual guidance. Bring the tip of the electrode close to the soma of the cell you are interested, form a sela and then suck up the the cell up the electrode. For details of solutions and protocols see our publication below.
best wishes, Refik
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