Open AccessTHE USE OF SPECIFIC ANTIBODY IN ELECTRON MICROSCOPY
Author(s) -
Frank A. Pepe,
Henry Finck,
Howard Holtzer
Publication year - 1961
Publication title -
the journal of cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.414
H-Index - 380
eISSN - 1540-8140
pISSN - 0021-9525
DOI - 10.1083/jcb.11.3.533
Subject(s) - staining , electron microscope , biology , antibody , fluorescence microscope , microbiology and biotechnology , negative stain , microscopy , heavy meromyosin , immunogold labelling , antigen , biophysics , fluorescence , myosin , pathology , immunology , genetics , physics , quantum mechanics , optics , medicine
Antibody staining was observed in the electron microscope by means of untagged antibody and osmium fixation. The antibody was visualized as a change in morphology due to its deposition on the antigenic structures. Glycerinated chicken breast muscle was stained with antimyosin, anti-H-meromyosin, and antiactin. The staining patterns obtained by electron microscopy were consistent with those previously demonstrated by fluorescence microscopy. A second method was used for confirmation of antibody staining. This consisted of extraction of unstained portions of the sarcomere with 0.6 M potassium iodide, 10-4 M adenosine triphosphate solution. Stained regions of the sarcomere remained intact because of insolubility of the combined antigen and antibody.
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