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Immunoelectron microscopic localization of elastic tissue components in archival tissue samples
Author(s) -
Fanning Joseph C.,
White Jacinta F.,
Polewski Roman,
Cleary Edward G.
Publication year - 1991
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1991.tb03146.x
Subject(s) - immunoelectron microscopy , materials science , electron microscope , pathology , chemistry , immunohistochemistry , medicine , physics , optics
SUMMARY Tissue samples that have been stored for many years, in different media and under a variety of conditions, have been examined by modern techniques of immunoelectron microscopy, using antibodies against elastic tissue components. A range of post‐embedding restorative procedures has been identified, which will allow reliable immunolocalization of antibodies against the elastic tissue component of such specimens. These methods have been applied successfully to autopsy‐derived material, fixed in buffered formaldehyde, to archival material stored frozen at −70 or −20·C, to specimens fixed for electron microscopy and stored for many years in buffer, and even to archival material from formaldehyde‐fixed, paraffin‐embedded blocks, reprocessed for electron microscopic examination. The successful restorative methods included pre‐treatment of the sections with 6***M guanidine hydrochloride, or 1M Tris/saline, each containing 100 ***mM dithiothreitol (a reducing agent) followed by alkylation with 220 mM iodoacetamide. The application of these techniques allowed reliable study of elastic tissue antibody distributions in archival tissues that could not be obtained again, as well as comparative studies with tissues processed many years previously.