Hostname: page-component-84b7d79bbc-l82ql Total loading time: 0 Render date: 2024-07-27T19:29:51.272Z Has data issue: false hasContentIssue false

A New Pre-Embedding Immunogold Method That Permits to Obtain a Very High Signal with a Very Good Ultrastucture

Published online by Cambridge University Press:  02 July 2020

G. Grondin
Affiliation:
Microscopy Service, Department of Biology, Faculty of Science, Université De Sherbrooke, Sherbrooke, Québec, CanadaJ1k-2r1.
A.R. Bcaudoin
Affiliation:
Microscopy Service, Department of Biology, Faculty of Science, Université De Sherbrooke, Sherbrooke, Québec, CanadaJ1k-2r1.
Get access

Abstract

The ultimate goal of the immunocytochemistry is to get the best signal with the lowest background while preserving ultrastructure. Unfortunaly too often the experimentator faces a compromise between density of the signal and ultrastructure preservation (1). with the advent of hight resolution confocal light microscopy, two photon imaging and other new technologies there is a need to provide the best immunocytochemical complementary results with the high resolution of the electron microscope. But often for exemple the high level of labelling observed by immunofluorescence microscopy is not matched by immunocytochemistry (2). One approach to obtain comparable levels of labelling is to use comparable protocols. We propose here such a new immunogold method. to illustrate our point we used a very convenient biological material, the adherent cells grown on 35 mm plastic culture dishes. The following procedure was applied : Confluent endothelial cells were fixed in situ for 3 hours with the following freshly prepared and filtered solution : 1 % 1-lysine, 4% paraformaldehyde, 0.04% glutaraldehyde, 0.25% sodium metaperiodate in 0.04M sodium cacodylate buffer, pH 7.4.

Type
Labeling for Microscopy and Correlative Microscopy (Organized By R. Albrecht)
Copyright
Copyright © Microscopy Society of America 2001

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

references

1-Skepper, J.N.Journal of Microscopy 199(2000)136.CrossRefGoogle Scholar
2-Webster, P.Microscopy today 9(2000)2834.CrossRefGoogle Scholar
3J., -Sevigny, et al., BBA 1334(1997) 7378.Google Scholar
4J.F., -HainfeldProc.57th.Mtg Micros. Soc.Am. Springer Berlin Heidelberg New York (1999)486487.Google Scholar
5-Geuze, H.J.Trends in Cell Biology 9(1999)9299.CrossRefGoogle Scholar