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Electron Tomography of Frozen-Hydrated Samples

Published online by Cambridge University Press:  02 July 2020

M. Marko ,
C.-E. Hsieh ,
B.K. Rath ,
C.A. Mannella  and
B.F. McEwen
M. Marko
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, Empire State Plaza, Albany, NY 12201-0509
C.-E. Hsieh
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, Empire State Plaza, Albany, NY 12201-0509
B.K. Rath
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, Empire State Plaza, Albany, NY 12201-0509
C.A. Mannella
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, Empire State Plaza, Albany, NY 12201-0509
B.F. McEwen
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, Empire State Plaza, Albany, NY 12201-0509
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Extract

Cryo-electron tomography offers a means of obtaining the 3-D ultrastructure of specimens that have not been chemically fixed or stained. The technique is still under development, but it has already been applied to several biological specimens including prokaryotic cells, centrioles, sperm axonemes, mitochondria, spindle-pole bodies, lipid vesicles, isolated skeletal-muscle triad junctions, chromatin fragments, actin bundles, and macromolecules.

Electron tomography itself is a nearly mature technology. It is most often used with conventional plastic sections 100- 1000nm thick. The specimen is imaged in the TEM, using a tilt stage to record a series of projection images over a large angular range. Using image-processing techniques, the tilt images are back-projected to form a reconstructed volume of the specimen. The reconstruction is a 3-D array of pixels or volume elements (voxels), and can be sliced in any direction to form a series of 2-D images.

Type
Technologists Forum: Cryo Microscopy
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 310 - 311
Copyright
Copyright © Microscopy Society of America

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