General Research Topics > Neuroscience

Golgi stain together with antibody stain? Sparse neuronal label?

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Dax42:
Dear all,

I was wondering whether I could somehow combine a golgi stain with an antibody stain, so do one of them first and then do the other one afterwards... Haven't really found anyone doing this, does one method prevent the other?

I basically need to be able to see three things: the cell body, the axon as it comes out of the cell body (say the first 50-100um) and an antibody label that overlaps with the axon. In order to be able to follow the structures properly, the cell labelling needs to be fairly sparse. I'm hoping to look at neurons in hippocampal and/or cortical slices AFTER the tissue has been fixed, which unfortunately rules out most sparse labelling techniques.

Any ideas would be very welcome!

gula:
http://eprints.uniss.it/5915/1/Spiga_S_Simultaneous_golgi-cox_and_immunofluorescence.pdf
this article deals with simultanous staining with Golgi and IF. As far as it works for IF, IHC will also do I think.
Have you made some trials?

Silver precipitates are not easily removed from tissue. I have no practical experience. just out of theory, I would stain Golgi first.
If antibody retrieval is needed I would compare doing it before or after Golgi and then proceed the IHC. Boiling in hot acid or basic solutions could influence the silver deposits. Perhaps protease digestion would be the better choice for AR.
basic more than acid.
Just try.

Gula

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