Ryan:
  Brain and CNS tissues in general are very tricky to process because all the lipid contents of the white matter and the fact that it is distributed sometimes unevenly through the tissue and that tissues from different areas have different rations of white/gray matter.
  What you are describing is caused by poor infiltration and that has nothing to do with the instrument you use, or the "freshness" of your reagents, but with the protocol and the time the CNS tissues are left in each step.  You have to have a protocol that assures the correct infiltration of the white matter to assure that it will not separate either when sectioning or staining (as is happening to you).
  Once you get to a blook poorly or unevenly infiltrated it is irrelevant what you try to do with the section itself. The tissue has already been poorly infiltrated and that is essentially irreversible, not matter how many anecdotal experiences to the contrary may lead you to believe otherwise.
  You have to develop a protocol suitable for infiltrating white matter, and this will also be suitable for the gary matter areas, with more cells and much less lipids.
  If at all possible discard your blocks and find a good protocol to work with. At the end will be more straight forward and will serve you for years to come and will avoid all the frustrations you are going through now.
  René J.
   
  
Ryan Dominique Salazar  wrote:
  
 



       
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