[Histonet] Some questions about cryosectioning
I have some questions about cryosectioning.
We use the following protocol for brain tissue of xenopus (a kind of frog)
a.. Bouinn fixation by perfusion, one night postfixation
b.. Wash in alc 70%, washing away picricacid
d.. section and mount on glass.
Just a normal routine, I think. I am used to do everything in one week, I begin with the perfusion and at the end of the week I have my sections stained.
Now the question: because I have to do many tissue's I cannot handle it in one week. The immunostaining has to be done for all tissue's at the same time etc..
a.. What is the best step to leave the tissue in for a while?
Bouinn, alc 70%, sucrose, and still have good immunostainings and good sectioning.
What is for each different step the maximum time, does it mather anyway?
b.. Already mounted on the slides , how long can you leave them (RT or 4C) on the slide and still have good immunoreactivity.
Dept. Cellular Animal Physiology
University of Nijmegen
6525 ED Nijmegen
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