X-Message-Number: 11557
Date: Thu, 15 Apr 1999 09:12:23 -0700 (PDT)
From: Doug Skrecky <>
Subject: ethylene glycol dehydration

Authors
  Stirling JW.
Institution
  Department of Pathology, Flinders Medical Centre, Bedford
  Park, Australia.
Title
  Unfixed tissue for
  electron immunocytochemistry: a simple preparation method
  for colloidal gold localization of sensitive epitopes using
  ethanediol dehydration [published erratum appears in Histochem J 1992
  Oct;24(10):757].
Source
  Histochemical Journal.  24(4):190-206, 1992 Apr.
Abstract
  A quick, simple protocol is described for the preparation of
  tissue for electron
  immunocytochemistry without the use of fixatives or deleterious solvents.
  Fresh, normal human colon was rapidly dehydrated in ethanediol (ethylene
  glycol) then embedded directly in low-acid glycol methacrylate. Using both
  mono- and polyclonal antibodies, in conjunction with colloidal gold probes, a
  range of intra- and extracellular epitopes were localized; these epitopes
  included lysozyme, chromogranin, desmin and collagen IV. Overall, the
  tissue compared well with material fixed in glutaraldehyde,
  partially dehydrated and embedded in LR White acrylic resin. Ultrastructural
  detail was good and was further enhanced, without affecting probe density and
  epitope localization, by the addition of 1% tannic acid or 1% uranyl acetate
  to the dehydrant. The technique is applicable to a wide range of
  tissues, allowing excellent antigen retention which might
  prove useful for the immunolocalization of sensitive
  epitopes.

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