Re: [Histonet] sections falling off

From:Rene J Buesa

If they were cryosectioned she did not have to go through xylene or graded alcohols to "dewax and hydrate" since there was no dehydration or wax infiltration to begin with.
  Cryosectioned sections just need to wash away the medium used to cryosection (OCT perhaps?) and just stain with the aqueous solutions.
  Later they can de dehydrated and cleared if a permanent mount is desired.
  Try to float the sections in a water bath and pick them up and don't try again to use xylene or alcohols.
  Probably she was following a procedure for paraffin embedded tissue that should have been adapted to frozen sections and it was not.
  René J.

Olek Michalski  wrote:
Dear Histonetters,

a friend of mine just faced a massive sections falling off. She is doing 
Nissl staining in mouse brain (brains perfused with formalin, postfixed 3 
days, and cryosectioned) on poly-L-lysined slides. She just managed to 
pass trough xylene and decreasing grades of alcohol to water and the 
sections started to detach. We were trying to reattach them but it went 
out that sections are not flat any more. It seems like the tissue is 
rehydrated unevenly, but keeping it in water for hours didn't work. Is 
there any way to spread these sections not damaging them at the same time? 
She is likely to rescue this sections even if some work is needed.

Best regards
Olek Michalski
Laboratory of Neurobiology
of Development and Evolution

Nencki Institute of Experimental Biology
ul. Pasteura 3, 02-093 Warszawa, Poland
Tel. +48 22 5892268, Fax +48 22 8225342

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