PMID: 6167611Jul 1, 1981Paper

Importance of fixation in immunohistochemistry: use of formaldehyde solutions at variable pH for the localization of tyrosine hydroxylase

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
A BerodJ F Pujol

Abstract

Adequate fixative in immunohistochemistry requires not only a rapid and total immobilization of the antigen, but also a sufficient preservation of its immunoreactivity and maintenance of its accessibility to the immunochemical reagents for localization. Thus, the optimal fixation condition for a specific antigen necessitates a compromise between these opposing variables and can be determined by the preparation of a series of tissues with a progressively increasing degree of fixation. Unless the results of localization using such a series is available, one must be satisfied with adequate but less than optimal results. In the present study, this principle is demonstrated using the localization of tyrosine hydroxylase in the dopaminergic system with formaldehyde as the fixative. The rate and degree of fixation with formaldehyde was shown to be highly pH dependent. By perfusing the tissue with formaldehyde at pH 6.5 (where the rate of fixation is extremely slow) it is possible to rapidly distribute the fixative homogeneously into the tissue. By suddenly changing to a formaldehyde perfusate of higher pH, the cross-linking reaction is rapidly increased. This two-step fixation procedure provides a means of obtaining a rapid and unifor...Continue Reading

Citations

Aug 14, 1998·The Journal of Comparative Neurology·C L Dolorfo, D G Amaral
Oct 8, 1998·The Journal of Comparative Neurology·A Malick, R Burstein
Sep 12, 2000·The Journal of Comparative Neurology·M U NaikT C Sacktor
Jul 1, 1990·Journal of Electron Microscopy Technique·N B SlepeckyL Cefaratti
Aug 1, 1990·Molecular Reproduction and Development·J A AmatU K Schubart
Sep 1, 1994·Cell and Tissue Research·L J Andries, D L Brutsaert
Apr 1, 1985·The Histochemical Journal·D Hopwood
Jun 5, 2008·Histochemistry and Cell Biology·Axel SteinkeEsther Asan
Apr 5, 2011·Brain Structure & Function·Saturnino SpigaMarco Diana
Oct 3, 1983·Brain Research·J E PintarX O Breakefield
Jul 8, 1985·Brain Research·B D BossW M Cowan
Aug 16, 1988·Brain Research·K D Cliffer, G J Giesler

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