[Histonet] croystat sectioning

[Edmondson David (RBV) NHS Christie Tr]

Kathleen
Like the Bright man says, that works for stopping the section rolling up as
it's cut, catch the leading edge as it comes off the blade and hold until
the section is there in all its glory. You have been doing this for years.  
But do you mean that it is rolling up when places on the PBS?
If so then all I have as an idea is to put them face down on the PBS. Rather
than face-up
Dave
Manchester. UK

-----Original Message-----
From: Kathleen A Spencer [mailto:kspencer <@t> utmem.edu]
Sent: 08 March 2004 15:47
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] croystat sectioning


I have a  problem with cryostat sectioning that is driving me crazy. I hope
someone out there can help because we have exhausted our brains coming up
with solutions that have not helped. I am sectioning perfused rat brain on a
cryostat, 20u thick, for IHC on floating sections. I have been doing this
successfully for years.
Now the problem is that they are curling up into tight little rolls in the
PBS, which make them impossible to stain. They even curl as soon as they
come off the knife edge, so I can't even put them on slides. 
We have: tried several different formulas for the fix, 3 different people
doing the perfusion, 3 different kinds of PBS, warm PBS, cold PBS, all
possible knife angles, different knives, different side of the knife, made
the block colder, or warmer, thicker sections, thinner sections,
cut fast, cut slow, NOTHING HELPS!
When we cut fresh frozen brain to put on slides we do not have this problem.
So, naturally we think it is the fix, or the method, or the person doing the
perfusion. So, I had the expert do a perfusion and it still happened. At
this point I just wanted to go out and get drunk and I am not a drinking
person. 
Can anyone help? All advice is appreciated.

Kathleen Spencer
Lab Manager
UTHSC


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