Hostname: page-component-5c6d5d7d68-pkt8n Total loading time: 0 Render date: 2024-08-06T22:32:08.806Z Has data issue: false hasContentIssue false

The Power of Freeze-Fracture Technique in Electron Microscopy

Published online by Cambridge University Press:  02 July 2020

B. Papahadjopoulos-Sternberg*
Affiliation:
Dept. of Microbiology, School of Dentistry, University of the Pacific, San Francisco, CA94115
Get access

Abstract

In the early 1960s, concerns about artifacts in preparing biological material for electron microscopy led to a new technique whereby samples are rapidly frozen, fractured under high vacuum, the fractured surfaces shadowed and replicated with a thin metal-carbon coat, and the cleaned replica examined in a transmission electron microscope. Pioneered by Moor and Muhlethaler subcellular structures are revealed with extraordinary three-dimensional clarity at near-molecular resolution. Furthermore, it was observed and proven at model systems as well as biological membranes that lipid bilayers split along their hydrophobic interior during freeze-fracture procedure. Therefore, freeze-fracture electron microscopy (FFEM) has the unique advantage of accessing the hydrophobic interior of biological (FIG.l, 5 and 6) as well as artificial bilayers (FIG. 2, 4, 5, and 6). Here it permits study of pattern generated by intrinsic proteins as well as lipids (FIG. 1 and 2).

Type
Recent Techniques for the Fixation and Staining of Biological Samples (Organized by M. Sanders and K. McDonald)
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.Moor, H. and Mühlethaler, K.. J. Cell Biol. 17 (1963) 609.CrossRefGoogle Scholar
2.Branton, D.. Proc. Natl. Acad. Sci. U. S. A. 55 (1966) 1048.Google Scholar
3.Deamer, D.W. and Branton, D.. Science 158 (1967) 655.CrossRefGoogle Scholar
4.Pinto da Silva, P. and Branton, D.. J. Cell Biol. 45 (1970) 598.CrossRefGoogle Scholar
5.Sternberg, B. et al, J. Struc. Biol. 110 (1993) 196.CrossRefGoogle Scholar
6.Dempsey, C.E. and Sternberg, B.. Biochim. Biophys. Acta 1061 (1991) 175.CrossRefGoogle Scholar
7.Sternberg, B.. In Liposome Technology, CRC Press (1992) 363.Google Scholar
8.Sternberg, B. et al, Nature 378 (1995) 21.CrossRefGoogle Scholar
9.Sternberg, B. et al., FEBS Lett. 356 (1994) 361.CrossRefGoogle Scholar
10.Sternberg, B. et al., Biochim. Biophys. Acta 1375 (1998) 23.CrossRefGoogle Scholar
11.Sternberg, B., J. Liposome Res. 6(1996) 515.CrossRefGoogle Scholar