RE: H&E problems

From:"Bennett, Catherine (Katie)" <>

Energy Beam Sciences has excellent information on methodologies for histo
staining of plastic sections on their website.  Go to:

My first thought as to the possible reasons for your wrinkle problems might
be due to over-dehydration if you are working with GMA embedded samples.
GMA samples are not fully dehydrated when processed as the highest
concentration of Ethanol is usually 95%.  If you are staining using reagents
that have ethanol solutions close to that or stronger, and/or dehydrating
the slides fully before coverslipping, you may be drying things out too much
and causing the cracking and wrinkling in the plastic.

I'll be interested to hear how you solve this.  Keep us posted.

Catherine "Katie" Bresee Bennett
Sr. Research Technologist
Lovelace Respiratory Research Institute
Albuquerque, New Mexico

> -----Original Message-----
> From:	Andrea Voogt []
> Sent:	Wednesday, February 28, 2001 6:24 PM
> To:
> Subject:	H&E problems
> Hi all,
> I've been staining plastic-embedded sections with H&E and running into a
> few problems.  First of all, on most of the slides the sections (which
> weren't wrinkled when I began) come out with lots of small wrinkles
> throughout the plastic - including the tissue.  I think this might have
> something to do with either the length of staining in eosin (I've been
> using 8 minutes) or the 1-second rinse in 95% EtOH, because I added 95%
> EtOH to my finished (but no good) slide and the wrinkling got worse.
> However, I don't know how to correct the problem.
> The other problem I've been having is that the eosin looks cracked on the
> tissue - sort of like dried earth.  This creates a problem in that the
> "cracks" are easily mistaken for cell membranes, creating artificial
> boundaries between the cells.  But I've noticed that the cracks often go
> right through nuclei, so they can't be the actual cell membranes.
> I don't know if this makes a difference or not, but I've also noticed that
> my staining is turning out a lot darker than the last time I did it (a
> year ago).
> Thanks in advance for all your help!!
> Sincerely,
> Andrea Voogt
> Dordt College   

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