Tired tissue adhesive topic- sorry

From:Stephanie Moore <smoore@brandeis.edu>

Hello HistoWorld,

I have read LOTS and LOTS of HistoNet archives on the topic of "getting
the sections to stick"

I have been using gelatin/chrom subbed slides and have let sections dry
24-48 hours and have been having wrinkling of edges when processing the
tissue.  I section fixed brain on a cryostat.  I do nissl, cytochrome
oxidase, and acetylcholinesterase stains.  With the nissl stain I don't
have a problem baking the slides for better adhesion.  I don't feel that
I can do this with the enzymatic stains for fear of denaturing my
beloved enzymes.  I want to try VectaBond but upon receiving the fax on
the method noticed that for paraffin embedded slides you need to bake at
56c.  For 'fresh' frozen sections it just says allow to dry, fix,
process as usual. Well, I have FIXED frozen sections.  I have dried the
enzymatic stain candidate sections for 2 days and still have them coming
off gel/chrom subbed slides.  ESPECIALLY with AChE stain (quenching
peroxidase is rather effervescent and bubbles my sections right off). 
The fact that my sections go into a solution that is water based (i.e.
no phosphate buffer based solutions) makes them REALLY wrinkly, then
they fall off.  CO staining isn't too bad because they go into a
phosphate buffer solution, although they don't wrinkle, they tend to
detach around the edges or slide down the slide.  I have really had it
with getting NO DECENT RESULTS!

PLEASE I beg you to help me.

Sorry to bring this up again (esp. J.A. Kiernan...you have great
detailed posts!)
Stephanie Moore



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