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using dehydrated brains for cryosectioning - (Jan/20/2010 )

I am studying the development of brain in the Chicken ( Gallus gallus). I routinely make cryosections of the embryonic brain. For this, I fix freshly dissected brains in PFA, and then treat them with a sucrose gradient, till 30% sucrose, then embed them in OCT, and section them.

I do not have any embryos ready to be harvested, although I do have some embryonic brains that were dehydrated and stored in Methanol.

I wanted to know if I could rehydrate these brain tissue and run them through sucrose gradient to further them?
Does the earlier dehydration compromise the quality of the sections in any way?

My downstream application will be primarily in-situ hybridization.

Please help.



If your tissue was put through a methanol gradient, rather than moved directly from a hydrous solution into a high concentration of methanol, then the structure of your tissue should be largely unaffected. I used to do this with retinal tissue on occasion.


-Carlton H-