[Histonet] Some questions about cryosectioning

[Frouwke Kuijpers]

Hallo Everyone

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
  c.. sucrose
  d.. section and mount on glass.
  e.. immunostaining
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.
Thanks Frouwke

F.J.Kuijpers-Kwant
Dept. Cellular Animal Physiology
University of Nijmegen
Toernooiveld 1 
6525 ED Nijmegen
the Netherlands
frouwke <@t> sci.kun.nl