Immunocytochemical localization of intracellular antigens with SEM

Springer Science and Business Media LLC - Tập 12 Số 4 - Trang 435-447 - 1980
Nakane, P. K.1, Hartman, A. L.1
1Department of Pathology, University of Colorado, School of Medicine, Denver, USA

Tóm tắt

A method for the localization of intracellular antigens with a scanning electron microscope using peroxidase-labelling antibodies is described. A search for a hydrogen donor which may be deposited at the sites of antigen by enzymatic action and emit secondary electrons or generate backscatter electrons was made. It was found that when 4-chloro-1-naphthol was used, the peroxidase deposited reaction product which resulted in a strong secondary electron emission at the site of antigen. With this method, the presence of luteinizing hormone in secretion granules and other cytoplasmic structures of gonadotropic cells was demonstrated. The level of detection of intracellular antigens with this method is not as high as that detectable with light microscopical examination of the same specimens, that is, more reaction product at the site of antigen is required to be detectable with scanning electron microscopy than with light microscopy. In spite of the lack of high sensitivity, the intracellular antigens may be localized with the method described.

Tài liệu tham khảo

citation_title=Backscatter electron imaging of endogenous and exogenous peroxidase activity in rat bone marrow; citation_inbook_title=Proceedings of the Ninth Annual SEM Symposium; citation_publication_date=1976; citation_pages=177-178; citation_id=CR1; citation_author=R. P. Becker; citation_author=P. P. H. Bruyn; citation_publisher=IITRI citation_title=Visualization of subsurface structures in cells and tissues by backscattered electron imaging; citation_inbook_title=Proceedings of the Twelfth Annual SEM Symposium; citation_publication_date=1979; citation_pages=835-870; citation_id=CR2; citation_author=R. P. Becker; citation_author=M. Sogard; citation_publisher=IITRI citation_journal_title=Histochem. J.; citation_title=A critique of the contributions of immunoperoxidase cytochemistry to our understanding of pituitary cell function; as illustrated by studies of gonadotropes, corticotropes and endogenous pituitary GnRH and TRH; citation_author=G. V. Childs, D. G. Ellison; citation_volume=12; citation_publication_date=1980; citation_pages=405-18; citation_id=CR3 citation_journal_title=J. Microscopy; citation_title=Staining of biological material for the scanning electron microscope; citation_author=K. E. Carr, J. McGradey; citation_volume=100; citation_publication_date=1974; citation_pages=323-30; citation_id=CR4 citation_title=Scanning electron microscopy of circulating human T lymphocytes; citation_inbook_title=Proceedings of the Electron Microscopy Society of America Thirty-Seventh Annual Meeting; citation_publication_date=1979; citation_pages=92-93; citation_id=CR5; citation_author=E. Harven; citation_author=D. Pla; citation_author=N. Lampen; citation_publisher=Claitor's Publishing Division citation_journal_title=J. Histochem. Cytochem.; citation_title=The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique; citation_author=R. C. Graham, M. J. Karnovsky; citation_volume=14; citation_publication_date=1966; citation_pages=291-302; citation_id=CR6 citation_journal_title=J. exp. Med.; citation_title=Scanning electron microscopy of tobacco mosaic virus labeled lymphocyte surface antigens; citation_author=U. Hämmerling, A. Polliack, N. Lampen, M. Sabety, E. Harven; citation_volume=141; citation_publication_date=1975; citation_pages=518-23; citation_id=CR7 citation_journal_title=Histochem. J.; citation_title=A new specific, sensitive and non-carcinogenic reagent for the demonstration of horseradishperoxidase; citation_author=J. S. Fanker, P. E. Yates, C. B. Metz, A. Rustioni; citation_volume=9; citation_publication_date=1977; citation_pages=789-92; citation_id=CR8 citation_journal_title=Arch. Hist. Jap.; citation_title=Ferritin as a surface marker for immuno-scanning electron microscopy. Observation of individual ferritin particles on erythrocytes; citation_author=A. Hattori, Y. Matsukura, S. Ito, T. Fugita, J. Tokunaga; citation_volume=39; citation_publication_date=1976; citation_pages=105-15; citation_id=CR9 citation_journal_title=J. Histochem. Cytochem.; citation_title=Periodatelysin-pparaformaldehyde fixative: a new fixative for immunoelectron microscopy; citation_author=I. W. Mclean, P. K. Nakane; citation_volume=22; citation_publication_date=1974; citation_pages=1077-83; citation_id=CR10 citation_title=Detection of embryonic surface antigens using the SEM.; citation_inbook_title=Proceedings of the Eleventh Annual SEM Symposium; citation_publication_date=1978; citation_pages=893-898; citation_id=CR11; citation_author=M. M. Miller; citation_author=R. L. Teplitz; citation_author=O. Jahari; citation_publisher=IITRI citation_journal_title=J. Cell Biol.; citation_title=New immunolatex spheres: visual markers of antigens on lymphocytes for scanning electron microscopy; citation_author=R. S. Monday, W. J. Dreyer, A. Rembaum, S. P. S. Yen; citation_volume=64; citation_publication_date=1975; citation_pages=75-88; citation_id=CR12 citation_title=Localization of hormones with the peroxidase-labeled antibody method; citation_inbook_title=Methods in Enzymology; citation_publication_date=1975; citation_pages=133-144; citation_id=CR13; citation_author=P. K. Nakane; citation_publisher=Academic Press citation_title=Utilization of high resolution spherical marker for labeling of virus antigens at the cell membrane in conventional scanning electron microscopy; citation_inbook_title=Proceedings of the Ninth Annual SEM Symposium; citation_publication_date=1976; citation_pages=75-83; citation_id=CR14; citation_author=K. R. Peters; citation_author=H. H. Gschwender; citation_author=W. Haller; citation_author=G. Rutter; citation_publisher=IITRI citation_journal_title=Lab. Invest.; citation_title=Antigens of epithelial basement membranes of mouse, rat and man. A study utilizing enzyme-labeled antibody; citation_author=G. B. Pierce, P. K. Nakane; citation_volume=17; citation_publication_date=1967; citation_pages=499-514; citation_id=CR15 citation_journal_title=J. Virol.; citation_title=Viral identification by scanning electron microscopy of preparations stained with fluorescein labeled antibody; citation_author=E. L. Springer, J. L. Riggs, A. J. Hackett; citation_volume=14; citation_publication_date=1974; citation_pages=1623-6; citation_id=CR16 citation_journal_title=Nature Lond.; citation_title=Fixation of ejaculated spermatozoa for electron microscopy; citation_author=M. Stefanini, C. Martino, L. Zamboni; citation_volume=216; citation_publication_date=1967; citation_pages=173-4; citation_id=CR17 citation_journal_title=Histochem. J.; citation_title=Immunochemistry on ultrathin frozen sections; citation_author=K. T. Tokuyasu; citation_volume=12; citation_publication_date=1980; citation_pages=381-403; citation_id=CR18