Recent advances in freeze-fracture electron microscopy: the replica immunolabeling technique

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Author: Robenek, H
Author: Severs, NJ
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Methods Article

Recent advances in freeze-fracture electron microscopy: the replica immunolabeling technique

Horst Robenek¹^* 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.

Figure 6 Enlarged

Fig. 6:

(A) Lipid droplet situated in a cup formed from ER membranes. Both ER membranes are visible (seen in P face (PF) and E face (EF)) view following the contour of the lipid droplet. The lipid droplet has been convexly fractured, and lies adjacent to and not within both ER membranes exposed. This example is from an unlabeled (standard freeze-fracture) preparation. (B, C) Similar views to (A), but with labeling for adipophilin using the FRIL technique. Abundant gold label is visible on the ER membranes (P face, PF) immediately adjacent to the lipid droplet. (D) Lipid droplet seen in cross fracture and in (E) concave fracture. FRIL demonstrates abundant labeling for adipophilin in the outer phospholipid monolayers surrounding the lipid droplets (P face, PF) exposed in these views. Bars: 0.2 µm

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