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Approaches for High Resolution Cryo-Electron Tomography of Biological Specimens

Published online by Cambridge University Press:  02 July 2020

D.A. Agard ,
M.B. Braunfeld ,
Hans Chen ,
Rebecca McQuitty  and
John Sedat
D.A. Agard
Affiliation:
The Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, CA94143-0448.
M.B. Braunfeld
Affiliation:
The Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, CA94143-0448.
Hans Chen
Affiliation:
The Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, CA94143-0448.
Rebecca McQuitty
Affiliation:
The Howard Hughes Medical Institute, University of California at San Francisco, San Francisco, CA94143-0448.
John Sedat
Affiliation:
Department of Biochemistry and Biophysics and University of California at San Francisco, San Francisco, CA94143-0448.
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Extract

Electron tomography is a powerful tool for elucidating the three-dimensional architecture of large biological complexes and subcellular organelles. Use of intermediate voltage electron microscopes extended the technique by providing the means to examine very large and non-symmetrical subcellular organelles, at resolutions beyond what would be possible using light microscopy. Recent studies using electron tomography on a variety cellular organelles and assemblies such as centrosomes (Moritz et al.,1995a,b), kinetochores (McEwen, 1993) and chromatin (Woodcock, 1994), have clearly demonstrated the power of this method for obtaining 3D structural information on non-symmetric cell components. When combined with biochemical and molecular observations, these 3D reconstructions have provided significant new insights into biological function.

Although the information that tomography provides is unique, its use as a general tool in the biological community has been limited due to the complexities involved in data collection and processing.

We are simultaneously trying to make this approach accessible through automation as well as trying to extend the resolution and accuracy of the reconstructions. Significant, has been the use of low-dose cryo-electron microscopic automated data collection methods.

Type
Computational Advances and Enabling Technologies For 3D Microscopies in Biology
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 1111 - 1112
Copyright
Copyright © Microscopy Society of America 1997

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References

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