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Effect of tissue fixation on anti-bromodeoxyuridine immunohistochemistry
B Schutte, MM Reynders, FT Bosman and GH Blijham
Department of Internal Medicine, University of Limburg, Maastricht, The Netherlands.
We investigated the influence of various fixatives on monoclonal anti- BrdUrd antibody binding of BrdUrd-substituted DNA in tissue sections of routinely processed mouse small intestine after in vivo administration of BrdUrd. For denaturing fixatives such as ethanol or Carnoy's fluid, a standard denaturation protocol showed specific crypt cell labeling. With cross-linking agents such as formalin and glutaraldehyde, a remarkable increase in staining intensity was obtained after tissue digestion with pepsin before acid denaturation. The optimal pepsin concentration was determined for maximal immunoreactivity combined with acceptable morphology.
Volume 35,
Issue 11,
pp. 1343-1345,
11/01/1987
Copyright © 1987 by The Histochemical Society
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