Hostname: page-component-77c89778f8-sh8wx Total loading time: 0 Render date: 2024-07-18T21:25:34.959Z Has data issue: false hasContentIssue false

Cryosections for Immunocytochemistry

Published online by Cambridge University Press:  02 July 2020

Paul Webster*
Affiliation:
House Ear Institute, 2100 West Third Street, Los Angeles, CA90057.
Get access

Extract

Immunocytochemistry describes methods for visualization of specific intracellular molecules within cells. These methods use high affinity markers to specifically bind to target molecules in tissues or cells. For electron microscopy, the sites where these markers are bound are visualized using electron opaque probes. Successful immunocytochemistry depends upon immobilization of target molecules, retention of subcellular morphology, and upon total accessibility of affinity marker to the target molecules.

Accessibility to extracellular molecules is not usually a problem, but for intracellular molecules, labeling protocols must be able to preserve subcellular morphology and allow the affinity markers to gain access to their target molecules. Currently, the best way to gain access to the inside of cells for electron microscopy (TEM) is to cut thin sections through them. In this way, morphology is retained and affinity markers can be applied to the cut face of the cells.

Type
Low Temperature Methods for Immunolabeling of Cells and Tissues
Copyright
Copyright © Microscopy Society of America

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1. Tokuyasu, K. T.. J. Cell Biol. 57(1973)551.CrossRefGoogle Scholar

2. Köhler, S. et al, Science 275(1997)1485.CrossRefGoogle Scholar

3. Liou, W., et al., Histochem. Cell Biol. 106(1996)41.CrossRefGoogle Scholar

4. Webster, P.. Methods in Molecular Biology 117(1999)49.Google Scholar

5. Griffiths, G., in Revel, J.-P. et al, Eds., Proc. Congr. Specimen Preparation (1984)153.Google Scholar

6. Keller, G. A. et al., Proc. Natl. Acad. Sci. USA 81(1984)5744.CrossRefGoogle Scholar

7. Tokuyasu, K. T.. Histochem. J. 21(1989)163.CrossRefGoogle Scholar

8. Anderson, R. G. et al., Proc. Natl. Acad. Sci. USA 81 (1984)4838.CrossRefGoogle Scholar

9. Slotetal., J. W., J. Cell Biol. 113(1991)123.Google Scholar