[Histonet] Tape transfer sections and IHC

Gayle Callis gcallis <@t> montana.edu
Fri Apr 15 12:19:41 CDT 2005


The matrix is the polymer which has been polymerized by UV light.  It is 
more of a problem after IHC and using an aqueous mounting media.

Several things:

If your chromogen is DAB, just spend more time in alcohols and xylenes to 
dissolve away the polymer as this does not damage the DAB.  Xylene 
substitutes don't work well, so keep a supply of xylene around for this 

For AEC where you must use an aqueous mounting media, use Crystal Mount 
(from Fisher), and be very generous when using it.  Spread it carefully 
over the section and surrounding area so it really covers the section well 
before drying.  Dry according to product insert directions and then mount a 
permanent coverslip over the Crystal Mount.  The polymer is there but 
greatly diminished, less visible, or may seem to disappear all 
together.  It fills IN the "bubbly" looking polymer matrix.

Also, if you use 4X slides, the polymer left over is much worse since this 
is 4 times thicker than 1X slides.    For bone work we found the 1/2 X 
slides are just as good, but we do flash the UV twice.  We occasionally use 
1X slides, and never 4X anymore. It would have to be the totally impossible 
sample before we use 4X.

To do our IHC, I actually use a teflon coated razor blade to scrape away 
the polymer from around the section, then use a Vectro ImmEdge pen for the 
hydrophobic barrier.   The reagents stay in place so much better without 
all that polymer there.

At 10:37 AM 4/15/2005, you wrote:

>I had problems with a 'matrix' looking  background and Bernice said they
>were working on  a special mounting media. I know she checks the histonet
>so hopefully she will answer 

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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