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Ultrastructural localization of tubulin and actin in polyethylene glycol‐embedded rat seminiferous epithelium by immunogold staining
Author(s) -
Wolosewick J. J.,
Mey J.,
Meininger V.
Publication year - 1984
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1111/j.1768-322x.1984.tb00240.x
Subject(s) - immunogold labelling , biology , immunocytochemistry , microbiology and biotechnology , ultrastructure , polyethylene glycol , staining , immunofluorescence , cytoplasm , tunica albuginea (penis) , centriole , actin , tubulin , cytoskeleton , glutaraldehyde , immunolabeling , microtubule , pathology , anatomy , cell , antibody , immunohistochemistry , biochemistry , immunology , medicine , penis , genetics , endocrinology
The polyethylene glycol (PEG) method for immunofluorescence localization of cytoskeletal antigens has been extended to the ultrastructural level using glutaraldehyde‐fixed tissues and immunogold staining. Semithin sections of fixed tissue embedded in polyethylene glycol are divested of the PEG, exposed to purified antibodies (e.g., antiactin, antitubulin) and anti‐IgG‐colloidal gold. The sections may be processed by dehydration and critical‐point drying, or reembedment in hydrophilic substances. Tubulin is demonstrated in the mitotic spindles of dividing spermatogonia, manchettes, axonemes and centrioles of developing spermatids, and in the Sertoli cell cytoplasm; actin localization is demonstrated in the myoid cells of the tunica propria, and smooth muscle cells of arterioles in the interstitial tissue. The results demonstrate the applicability and versatility of PEG embedding for immunocytochemistry.