SpringerProtocols: Abstract: 8. Overview of Cell Fixatives and Cell Membrane Permeants

8. Overview of Cell Fixatives and Cell Membrane Permeants

By: Melissa A. Melan²

Abstract

Full Text

Download PDF (175K)

The localization of proteins and carbohydrates within cells and tissues with specific antibodies has long been proven to be a valuable technique. Immunolocalization procedures allow one to detect not only well-characterized cellular structures but also provide information about newly characterized proteins and carbohydrates. This chapter will review some of the advantages and drawbacks of common chemical fixation and permeabilization methods used for immunolocalization at the level of light microscopy.

Affiliation(s): (2) Biology Department Duquesne University, Pittsburgh, PA

Book Title: Immunocytochemical Methods and Protocols

Series: Methods in Molecular Biology | Volume: 115 | Pub. Date: Jan-15-1999 | Page Range: 45-55 | DOI: 10.1385/1-59259-213-9:45

Subject: Immunology

References

Download References

1.	Larsson, L. I. (1988) Immunocytochemistry:Theory and Practice. CRC, Boca Raton, FL.

2.	Wang, Y. L. (1989) Fluorescent analog cytochemistry: tracing functional protein components in living cells, in Methods in Cell Biology, vol. 29 (Taylor, D. L. and Wang, Y. L., eds.), Academic, San Diego, CA, pp. 1–12.

3.	Hyat, M. A. (1981) Fixation for Electron Microscopy. Academic, New York.

4.	Bowers, B. and M. Maser. (1988) Artifacts in fixation for transmission electron microscopy, in Artifacts in Biological Electron Microscopy (Crang, R. F. E. and Klomparens, K. L., eds.), Plenum, New York, pp. 13–42.

5.	Humason, G. L. (1967) Animal Tissue Techniques. W. H. Freeman, San Francisco.

6.	Melan, M. A. and Sluder, G. (1992) Redistribution and differential extraction of soluble proteins in permeabilized cultured cells: implications for immunofluorescence microscopy. J. Cell Sci. 101, 731–743.

7.	Huber, R., Deisenhofer, J., Coleman, P. M., Matsushima, M., and Palm, W. (1976) Crystallographic structure studies of an IgG molecule and an F_c fragment. Nature 264, 415–420.

8.	Poljak, R. J., Amzel, L. M., Avey, H. P., Becka, L. N., and Nissonoff, A. (1972) Structure of Fab&#X2019; New at 6Å resolution. Nature New Biol. 235, 137–140.

9.	Glauert, A. M. (1974) Fixation, dehydration and embedding of biological specimens, in Practical Methods in Electron Microscopy, vol. 3 (Glauert, A. M., ed.), North-Holland, Amsterdam, pp. 1–201.

10.	Goldenthal, K. L., Hedman, K., Chen, J. W., August, J. T., and Willingham, M. C. (1985) Postfixation detergent treatment for immunofluorescence suppresses localization of some integral membrane proteins. J. Histochem. Cytochem. 33, 813–820.

11.	Kiernan, J. A. (1981) Histological and Histochemical Methods. Theory and Practice. Pergamon, Oxford, UK.

12.	Ito, S. and Karnovsky, M. J. (1968) Formaldehyde-glutaraldehyde fixatives containing trinitro compounds. J. Cell Biol. 39, 168a.

13.	Newman, G. R., Jasani, B., and Williams, E. D. (1983) A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope. Histochem. J. 15, 543–555.

14.	Weber, K., Rathke, P. C., and Osborn, M. (1978) Cytoplasmic microtubular images in glutaraldehyde-fixed tissue culture cells viewed by electron microscopy and by immunofluorescence microscopy. Proc. Natl. Acad. Sci. USA 75, 1820–1824.

15.	Leonard, J. B. and Shepardson, S. P. (1994) A comparison of heating modes in rapid fixation techniques for electron microscopy. J. Histochem. Cytochem. 42, 383–391.

16.	Safiejko-Mroczka, B. and Bell, P. B. (1996) Bifunctional protein cross-linking reagents improve labeling of cytoskeletal proteins for qualitative and quantitative fluorescence microscopy. J. Histochem. Cytochem. 44, 641–656.

17.	Kendall, P. A., Polak, J. M., and Pearse, A. G. E. (1971) Carbodiimide fixation for immunohistochemistry: observations on the fixation of polypeptide hormones. Experientia 27, 1104–1106.

18.	Harris, P., Osborn, M., and Weber, K. (1980) Distribution of tubulin containing structures in the egg of the sea urchin Strongylocentrotus purpuratus from fertilization through first cleavage. J. Cell Biol. 84, 668–679.

19.	Assa, Y., Shany, S., Gestetner, B., Tencer, Y., Birk, Y., and Bondi, A. (1973) Interaction of alfalfa saponins with components of the erythrocyte membrane in hemolysis. Biochim. Biophys. Acta 307, 83–91.

20.	Seeman, P. (1967) Transient holes in the erythrocyte membrane during hypotonic hemolysis and stable holes in the membrane after lysis by saponin and lysolecithin. J. Cell Biol. 32, 55–70.

21.	Neugebauer, J. (1988) A Guide to the Properties and Uses of Detergents in Biology and Biochemistry. Calbiochem-Novabiochem Corp., 1La Jolla, CA.

22.	Ashani, Y. and Catravas, G. N. (1980) Highly reactive impurities in Triton X-100 and Brij 35: Partial characterization and removal. Anal. Biochem. 109, 55–62.

23.	Melan, M. A. and Cosgrove, D. J. (1988) Evidence against the involvement of ionically bound cell wall proteins in pea epicotyl growth. Plant Physiol. 86, 469–474.

24.	Wick, S. M., Seagull, R. W., Osborn, M., Weber, K., and Gunning, B. E. S. (1981) Immunofluorescence microscopy of organized microtubule arrays in structurally stabilized meristematic plant cells. J. Cell Biol. 89, 685–690.

Comments (Loading...)

Post comment/View all comments