[Histonet] mounting very thick specimen

[Lee & Peggy Wenk]

Years ago, I had to mount whole liver flukes, which were thick, but not as
thick as your 400 um (0.4 mm). Here's how we did it - see if it works for
you:

Take another glass slide, and carefully break it into little pieces, like
1-2 mm square pieces. (The safety officer in me had to say carefully, and
please use safety goggles.) Take 4 little pieces, and place them flat around
the outside of the tissue - far enough away so if the pieces move, they
won't scratch the tissue, but close enough so that they would be located
under the coverslip. 

Use mounting media, and put LOTS of drops of it on the slide and around the
pieces of broken glass. Thicker viscosity (such as slightly old mounting
media with some of the solvent evaporated out) works a little better, as it
won't run all over the slide and the counter before it sets/dries. Coverslip
by hand with a glass slide. Slow motion coverslipping helps, so the glass
pieces don't move and bubble won't form. 

Since a lot of mounting media is used, it can take several days to set.
Until set, the coverslip is very easy to move, and the mounting media will
ooze out all over the slide, counter, and fingers. To speed up drying, place
the coverslipped slide flat in a 60 degree oven for a couple of hours to
overnight, to allow mounting media to solidify. Set the glass slide on top
of a flat lid from a coplin jar, on the screwtop lip (the flat top is
resting down on the shelf in the oven). That way, if any mounting media did
escape and flow to the backside of the glass slide, the slide isn't "glued"
to the oven shelf. (Learned that one the hard way.)

Sometimes, not enough mounting media is used, and when dried, it has pulled
inward away from the edge of the coverslip. Just add some more drops of
mounting media on the slide, touching the edge of the coverslip. The drop(s)
should wick into the empty space(s). Place back into the oven for a couple
of hours to solidify.

The coverslip is now being held up away from the tissue by the thickness of
the pieces of broken glass. Let us know if this technique works for your
project.

Peggy A. Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sebastien
Thuault
Sent: Wednesday, June 07, 2006 6:37 PM
To: Histonet
Subject: [Histonet] mounting very thick specimen

Does anyone have tips to mount very thick specimen (400 um) on slides in
aqueous or resinous medium?

Thanks!

Sebastien Thuault, PhD
Center for Neurobiology and Behavior
Columbia University
PI Annex/Kolb Institute
1051 Riverside Drive
New York, NY 10032

Phone: +1-212-543-5037
Fax: +1-212-795-7997


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