I need to patch clamp neurons on a substrate that scatters light so I can't use DIC or phase contrast to visualize cells. If I fluorescently label the cells, I have problems with the substrate and the pipette. I'm looking for a non-toxic stain that will dye the cells a blue, green, or red color.
Hi Vanessa,
You could try Biocytin labeling
Check the attached protocol and the link below
http://neurobio.drexelmed.edu/GaoWeb/images.html
Did you try filling the pipettees with Alexa fluor 594 in your pipettes which seems to a standard method
http://neuronet.jp/pdf/O_095.pdf
Also Another possibility is to transfect your neurons with mcherry construct. that way you could visualize the neuronal cell soma and the neurites in their entirety
Vanessa, I was wondering if you found a solution to your problem, which is very similar to mine. The suggestions proposed by Karthik do not apply to my case.
In our case, after a lot of trying, the only thing that remotely colored the cells on our surface in a way we could see without using fluorescence was MTT. We used a normal staining and then instead of lysing and putting it on a plate reader we tried to do patch clamp. The cells were slightly pink so we could kind of see them. Unfortunately, when switching to patch clamp solutions the remaining MTT made a lot of precipitates that tended to clog the tips so in the end it wasn't a way to patching, but if you just need to find where on a substrate a cell is, it kind of worked.
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