[Histonet] Frozen section problems!!

[Adam M Gomez]

   We  are  wanting  to  section  peripheral  gang   cryostat. Our current procedure is as follows:
   Fixation:
   1)perfused with PBS/8% Para, ganglia tissue removed
   <   color   10 min.
   3)placed in 30% sucrose overnight (or until   4)rinsed in PBS/dH20
   5)embedded in OCT
   6)frozen rapidly in liquid nitrogen
   7)tissue  allowed  to  equilibrate  to  cryostat temp & sectio   cryostat (-18 C)
   8)sections   hour.
   9)Slides  are  dipped  in clearing agent (xylenes)   alcohol  (95%, 70%, 50%), dH2O, cresyl violet stain, dH2   95%, & xylenes...and coverslipped immediately.

   The problem we are having is that the tissue looks outstanding after    sectioning  as  long  as  it remains wet (i.e., when we take a look at
   tissue  d   we're losing ce   seems  that  the  dehydr   tissue  is  the culprit. We have tr   procedure but have yet to find a successf   the  tissue.   If  you have any suggestions p   problem has been detrimental to our work for the pa
   Thanks,

      Adam Gomez
   Research&nb   University of Nebraska at Om   Department of Psychology
   (402) 554-6028
   
   
References

   1. 3D"mailto:amgomez <@t> mail.unomaha.edu"