[Abstract]  [Full Text]  [PDF]

Horst Robenek^1* and Nicholas J. Severs²

¹ Department of Cell Biology and Ultrastructure Research, Leibniz-Institute for Arteriosclerosis Research. University of Münster, Domagkstr. 3D-48149 Münster. Germany.
² National Heart and Lung Institute, Imperial College London. U.K..

* To whom correspondence should be addressed: Horst Robenek, Department of Cell Biology and Ultrastructure Research, Leibniz-Institute for Arteriosclerosis Research. University of Münster, Domagkstr. 3D-48149 Münster. Germany. Phone: +49-251-83-56426. Fax: +49-251-83-52998. Email: robenek@uni-muenster.de

Biol. Proced. Online 2008;10:9-19. doi:10.1251/bpo138
Submitted: October 18, 2007; Accepted: December 12, 2007; Published: January 28, 2008.

Indexing terms: Freeze Fracturing; Microscopy, Immunoelectron; Immunogold Techniques.

Freeze-fracture electron microscopy is a technique for examining the ultrastructure of rapidly frozen biological samples by transmission electron microscopy. Of a range of approaches to freeze-fracture cytochemistry that have been developed and tried, the most successful is the technique termed freeze-fracture replica immunogold labeling (FRIL). In this technique, samples are frozen, fractured and replicated with platinum-carbon as in standard freeze fracture, and then carefully treated with sodium dodecylsulphate to remove all the biological material except a fine layer of molecules attached to the replica itself. Immunogold labeling of these molecules permits their distribution to be seen superimposed upon high resolution planar views of membrane structure. Examples of how this technique has contributed to our understanding of lipid droplet biogenesis and function are discussed.