Abstract
For high resolution labeling of influenza virus cell surface antigens on HeLa cells, an immunospecific marker is used with silica sphere cores of 13--14 nm average diameter. These markers are formed using commercially available silica sphere sols. Two other size ranges are available, 7--8 nm and 22--25 nm. The steps for chemical derivatization are described in detail. Amino and aldehyde functions are covalently introduced onto the sphere surface. Sols of these derivatized silica spheres (DSS) are physicochemically stable and therefore usable for years. Coupling of IgG to DSS followed by permeation chromatography on controlled pore glass results in size-defined immunospecific silica sphere markers (DSS-markers). Saturation labeling of cell surface antigens on HeLa cells on cover slips is obtained with the final sphere concentration of 10(14) DSS-marker/cm3 within 20 min. With usual protective conditions, the marker stability and labeling ability are preserved for months. The visibility and the fine structure of the DSS- marker on cell surfaces are shown by using transmission electron microscopy (TEM) with stereo replicas and ultrathin sections.
Full Text
The Full Text of this article is available as a PDF (3.1 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Biberfield P., Ghetie V., Sjöquist J. Demonstration and assaying of IgG antibodies in tissues and on cells by labeled staphylococcal protein A. J Immunol Methods. 1975 Jan;6(3):249–259. doi: 10.1016/0022-1759(75)90068-x. [DOI] [PubMed] [Google Scholar]
- Campbell D. H., Luescher E., Lerman L. S. Immunologic Adsorbents: I. Isolation of Antibody by Means of a Cellulose-Protein Antigen. Proc Natl Acad Sci U S A. 1951 Sep;37(9):575–578. doi: 10.1073/pnas.37.9.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Crone H. D., Dawson R. M. Residual anionic properties of a covalently substituted controlled-pore glass, glyceryl-CPG. J Chromatogr. 1976 Dec 22;129:91–96. doi: 10.1016/s0021-9673(00)87771-7. [DOI] [PubMed] [Google Scholar]
- Fuchs H., Bächi T. Scanning electron microscopical demonstration of respiratory syncytial virus antigens by immunological markers. J Ultrastruct Res. 1975 Jul;52(1):114–119. doi: 10.1016/s0022-5320(75)80026-8. [DOI] [PubMed] [Google Scholar]
- Gschwender H. H., Haller W., Hofschneider P. H. Large-scale preparations of viruses by steric chromatography on columns of controlled pore glass. Phi-X174, M13, M12, Q-beta and T4 bacteriophages. Biochim Biophys Acta. 1969 Oct 22;190(2):460–469. doi: 10.1016/0005-2787(69)90095-1. [DOI] [PubMed] [Google Scholar]
- Hiatt C. W., Shelokov A., Rosenthal E. J., Galimore J. M. Treatment of controlled pore glass with poly(ethylene oxide) to prevent adsorption of rabies virus. J Chromatogr. 1971 Apr 7;56(2):362–364. doi: 10.1016/s0021-9673(00)97825-7. [DOI] [PubMed] [Google Scholar]
- Kay M. M. Mechanism of removal of senescent cells by human macrophages in situ. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3521–3525. doi: 10.1073/pnas.72.9.3521. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Molday R. S., Dreyer W. J., Rembaum A., Yen S. P. Latex spheres as markers for studies of cell surface receptors by scanning electron microscopy. Nature. 1974 May 3;249(452):81–82. doi: 10.1038/249081a0. [DOI] [PubMed] [Google Scholar]
- Nemanic M. K., Carter D. P., Pitelka D. R., Wofsy L. Hapten-sandwich labeling. II. Immunospecific attachment of cell surface markers suitable for scanning electron microscopy. J Cell Biol. 1975 Feb;64(2):311–321. doi: 10.1083/jcb.64.2.311. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peters K. R. Stereo surface replicas of culture cells for high-resolution electron microscopy. J Ultrastruct Res. 1977 Oct;61(1):115–123. doi: 10.1016/s0022-5320(77)90010-7. [DOI] [PubMed] [Google Scholar]
- Rutter G., Mannweiler K. Movement of virus-induced antigens on the cell surface. Arch Gesamte Virusforsch. 1973;43(1):169–172. doi: 10.1007/BF01249361. [DOI] [PubMed] [Google Scholar]
- Smith S. B., Revel J. P. Mapping of concanavalin A binding sites on the surface of several cell types. Dev Biol. 1972 Mar;27(3):434–441. doi: 10.1016/0012-1606(72)90183-2. [DOI] [PubMed] [Google Scholar]
- Spurr A. R. A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res. 1969 Jan;26(1):31–43. doi: 10.1016/s0022-5320(69)90033-1. [DOI] [PubMed] [Google Scholar]