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From: | "Dr. Ian Montgomery" |
Erin,
Can't say
that I've ever noticed any difference in the staining characteristics
using the various clearing agents. The major difference is the cutting
characteristics. I often find that if it's alcohol/xylene the tissue can
be a bit brittle but if, for example, alcohol/chloroform/xylene the
tissue sections very easily. Now, if your only after a couple of sections
then alcohol/xylene is fine but if, like me, you have to cut 5um serial
sections through a 1cm block of something like bird ovary/ova then you
want easy sectioning through the whole block. In the past I tried the
various limolene based agents but the smell, even reduced smell, gave me
the boak so I just gave that route a miss. My field, well, by training
I'm a Histophysiologist but after recent downsizing in the University I'm
it as regards the all rounder in microscopy. L.M., E.M., S.E.M.,
L.S.C.M., name it and I'm your boy. If it crawls, flies, swims or gives
you nasty diseases I have to do it. Benefits, I'm developing my skills
daily or using techniques I learned as a boy. Downside, probably the same
everywhere, I'm 54 and nobody here to train and pass on the skills both
technical and academic. Sad, but in a few years I imagine they'll be
re-inventing the H&E. It happened with E.M. over the last 10-20
years, confocals came in and E.M's went out. The pendulum has swung again
and now I find a large number of scientist know absolutely nothing about
the ultrastructure of cells. Check the book shops and look for an atlas
of ultrastructure. The classic, Rodin's Histology is no longer in print.
Sad, but fortunately we still have Histonet, a jewel shining
brightly.
Oh, PhD
from this University that was 550 years old last year.
Ian.
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Date: Thu, 24 Jan 2002 11:14:31 -0500
To: ian.montgomery@bio.gla.ac.uk
From: Erin Ellison <eellison@lakeridgehealth.on.ca>
Subject:
>I still use chloroform, cedarwood oil etc., "real" clearing
agents and when the tissue has been thoroughly dehydrated, cleared and
embedded the resulting blocks are a dream.
Do you really use these? I would never dare to say this in today's world,
but I am trying to hang on tooth and nail to as many of the "old
ways" as possible. Luckily, in some parts of Canada it is still
possible to use real alcohol and xylene. But I haven't been in the biz
long enough to have seen what you are referring with the "real"
clearing agents. When you say the blocks are a dream, is it just cutting
or also the stained sections? If the latter, would you happen to have any
comparative photos you could e-mail me? If I'm not being rude
asking, what sort of biz are you in? Are you PhD, MD (yours or ours) or
both? What is your field? From the notes you drop occasionally, your work
sounds fascinating.
Kind regards.
Erin Ellison
=================================
Erin Ellison MD FRCP(C)
Histocytopathologist
Laboratory Medicine Program
Lakeridge Health Corporation,
1 Hospital Court, Oshawa ON L1G 2B9 Canada
Tel 1(905) 576-8711 x4497 Fax 1 (905) 721-4757
eellison@lakeridgehealth.on.ca
Dr. Ian Montgomery,
Microscopy Service Unit,
Graham Kerr Building,
Institute of Biomedical & Life Sciences,
University of Glasgow,
Glasgow,
G12 8QQ.
Tel: 0141 332 8855 Extn.6644.
e-mail: ian.montgomery@bio.gla.ac.uk
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