(Fwd) RE: Sections lifting off

From:Margaret Horne

Thanks everyone for the many suggestions; hope has now replaced
dispair. I have forwarded this reply to you all in case it is of use to
others.  It is packed with great ideas.
             Thanks again , Margaret

------- Forwarded message follows -------
Date sent:      	Fri, 28 Jun 2002 08:32:34 -0500
From:           	Jeff Gordon 
Subject:        	RE: Sections lifting off
To:             	mhorne@upei.ca

Margaret, we at Cell Marque incorporate one of the most aggressive
unmasking protocols used today, which is EDTA in the pressure cooker for
15 minutes at 120 degrees.  It is essential to have the slides prepared
correctly in order to maintain tissue adhesion on the slides during this
aggressive pretreatment.  There are labs across the world that use our
protocol with great success, including both clinical and research labs.
We run all of our tissue the same way, whether it's skin, breast, lung,
brain, muscle, pancreas, etc.

In order to ensure that the tissue stays on the slide, the answer just
lies in the simplest preparation methods.  We just use standard
positively charged slides, with no special coating or adhesion.

1. Cut no thicker than 4 microns. (3 is ideal)

2. Dry 2-4 hours at 58 C in oven (4 hours minimum for breast tissue or
other fatty tissue).  You can also dry overnight at 37 C.  Finally,
there are microwave methods of drying tissue, but the protocol depends
on your microwave and the number of slides.  Generally, a 1000W
microwave can dry up to 100 slides in about 3 minutes.  You should
examine each slide to make sure the paraffin has melted uniformly across
the tissue section.

3. Make sure you use positively charged slides.  If the tissue adhesion
problem coincides with the use of a newly received shipment of slides,
you may have a bad lot of slides.

4. When lifting the tissue on to the slide out of the water-bath, at no
point before or during this process should you touch the flat surface of
the slide.  Hold the slides by the edges or by the front end.

By following these simple guidelines, you should have consistently good
results with your heat retrieval.  If you have any questions, please
contact me at 1-800-665-7284.

Jeff Gordon
Cell Marque Corp.
------- End of forwarded message -------
Margaret Horne
Dept. of A&P,
Atlantic Veterinary College, U.P.E.I.
550 University Ave., Charlottetown,
P.E.I.,  C1A 4P3

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