Abstract
A hapten-sandwich procedure has been used for immunospecific labeling of cell surface antigens with markers visible by scanning electron microscopy. Antihapten antibody was used to link hapten-modified tobacco mosaic virus, bushy stunt virus, or hemocyanin to hapten- modified human erythrocytes. The antihapten antibody bridge was also used to link the hapten-virus marker to hapten-modified antibodies against mammary tumor virus on mouse mammary tumor cells, or against immunoglobulin receptors on mouse splenic lymphocytes. In all cases, labeling was highly specific. With this technique, it is possible to (a) compare morphological features of cells bearing differing cell surface antigens, and (b) examine the arrangement of specific antigenic sites on a cell surface or their distribution relative to membrane structures such as microvilli.
Full Text
The Full Text of this article is available as a PDF (3.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aoki T., Boyse E. A., Old L. J., De Harven E., Hämmerling U., Wood H. A. G (Gross) and H-2 cell-surface antigens: location on Gross leukemia cells by electron microscopy with visually labeled antibody. Proc Natl Acad Sci U S A. 1970 Mar;65(3):569–576. doi: 10.1073/pnas.65.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aoki T., Wood H. A., Old L. J., Boyse E. A., De Harven E., Lardis M. P., Stackpole C. W. Another visual marker of antibody for electron microscopy. Virology. 1971 Sep;45(3):858–862. doi: 10.1016/0042-6822(71)90214-5. [DOI] [PubMed] [Google Scholar]
- Blair P. B., Smoller C., Bonhag R. S., Weiss D. W. Immunology of the mouse mammary tumor virus (MTV). Electron microscopic studies of interaction between MTV and specific antibody in immunodiffusion. J Natl Cancer Inst. 1968 Jun;40(6):1325–1338. [PubMed] [Google Scholar]
- Corneil I., Wofsy L. Specific purification of equine anti-SII antibodies by precipitation with a hemocyanin-glucuronide conjugate. Immunochemistry. 1967 May;4(3):183–189. doi: 10.1016/0019-2791(67)90127-9. [DOI] [PubMed] [Google Scholar]
- Das N. K., Hosick H. L., Nandi S. Influence of seeding density on multicellular organization and nuclear events in cultures of normal and neoplastic mouse mammary epithelium. J Natl Cancer Inst. 1974 Mar;52(3):849–861. doi: 10.1093/jnci/52.3.849. [DOI] [PubMed] [Google Scholar]
- Lamm M. E., Koo G. C., Stackpole C. W., Hämmerling U. Hapten-conjugated antibodies and visual markers used to label cell-surface antigens for electron microscopy: an approach to double labeling. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3732–3736. doi: 10.1073/pnas.69.12.3732. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lin P. S., Cooper A. G., Wortis H. H. Scanning electron microscopy of human T-cell and B-cell rosettes. N Engl J Med. 1973 Sep 13;289(11):548–551. doi: 10.1056/NEJM197309132891102. [DOI] [PubMed] [Google Scholar]
- Lin P. S., Wallach D. F., Tsai S. Temperature-induced variations in the surface topology of cultured lymphocytes are revealed by scanning electron microscopy. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2492–2496. doi: 10.1073/pnas.70.9.2492. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Linthicum D. S., Sell S. Letter: A rabbit blood lymphocyte is both smooth and hairy. N Engl J Med. 1974 Jun 6;290(23):1327–1328. doi: 10.1056/NEJM197406062902323. [DOI] [PubMed] [Google Scholar]
- Nemanic M. K., Pitelka D. R. A scanning electron microscope study of the lactating mammary gland. J Cell Biol. 1971 Feb;48(2):410–415. doi: 10.1083/jcb.48.2.410. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Polliack A., Lampen N., Clarkson B. D., De Harven E., Bentwich Z., Siegal F. P., Kunkel H. G. Identification of human B and T lymphocytes by scanning electron microscopy. J Exp Med. 1973 Sep 1;138(3):607–624. doi: 10.1084/jem.138.3.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Porter K. R., Todaro G. J., Fonte V. A scanning electron microscope study of surface features of viral and spontaneous transformants of mouse Balb-3T3 cells. J Cell Biol. 1973 Dec;59(3):633–642. doi: 10.1083/jcb.59.3.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Truffa-Bachi P., Wofsy L. Specific separation of cells on affinity columns. Proc Natl Acad Sci U S A. 1970 Jul;66(3):685–692. doi: 10.1073/pnas.66.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilkinson J. M. Variation in the N-terminal sequence of heavy chains of immunoglobulin G from rabbits of different allotype. Biochem J. 1969 Apr;112(2):173–185. doi: 10.1042/bj1120173. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wofsy L., Burr B. The use of affinity chromatography for the specific purification of antibodies and antigens. J Immunol. 1969 Aug;103(2):380–382. [PubMed] [Google Scholar]
- Wolfsy L., Baker P. C., Thompson K., Goodman J., Kimura J., Henry C. Hapten-sandwich labeling. I. A general procedure for simultaneous labeling of multiple cell surface antigens for fluorescence and electron microscopy. J Exp Med. 1974 Aug 1;140(2):523–537. doi: 10.1084/jem.140.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]