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Electron Microscopic Tomography of Cellular Organelles: Chemical Fixation Vs. Cryo-Substitution of Rat-Liver Mitochondria

Published online by Cambridge University Press:  02 July 2020

C.A. Mannella ,
K. Buttle ,
K. Tessitore ,
B.K. Rath ,
C. Hsieh ,
D. D'Arcangelis  and
M. Marko
C.A. Mannella
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, New York State Department of Health,Empire State Plaza, Box 509, Albany, NY 12201-0509, and Department of Biomedical Sciences, School of Public Health, University at Albany (SUNY)
K. Buttle
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, New York State Department of Health,Empire State Plaza, Box 509, Albany, NY 12201-0509, and Department of Biomedical Sciences, School of Public Health, University at Albany (SUNY)
K. Tessitore
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, New York State Department of Health,Empire State Plaza, Box 509, Albany, NY 12201-0509, and Department of Biomedical Sciences, School of Public Health, University at Albany (SUNY)
B.K. Rath
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, New York State Department of Health,Empire State Plaza, Box 509, Albany, NY 12201-0509, and Department of Biomedical Sciences, School of Public Health, University at Albany (SUNY)
C. Hsieh
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, New York State Department of Health,Empire State Plaza, Box 509, Albany, NY 12201-0509, and Department of Biomedical Sciences, School of Public Health, University at Albany (SUNY)
D. D'Arcangelis
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, New York State Department of Health,Empire State Plaza, Box 509, Albany, NY 12201-0509, and Department of Biomedical Sciences, School of Public Health, University at Albany (SUNY)
M. Marko
Affiliation:
Biological Microscopy and Image Reconstruction Resource, Wadsworth Center, New York State Department of Health,Empire State Plaza, Box 509, Albany, NY 12201-0509, and Department of Biomedical Sciences, School of Public Health, University at Albany (SUNY)
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Extract

Electron microscopic tomography is proving to be a valuable tool for investigating the 3D structure and organization of cellular organelles. Important progress is being made in the application of the technique to frozen-hydrated material, but it is likely that success with thick specimens will be limited by the low contrast and beam sensitivity of naked biological material. Thus, optimizing procedures for fixing, embedding, staining, and selectively labelling cells for 3D electron microscopy remains a priority.

Tomography of chemically fixed and plastic-embedded rat-liver tissue and isolated mitochondria has shown that the cristae (the invaginations of the mitochondrial inner membrane) are pleiomorphic and connected to each other and to the surface of the inner membrane by tubular regions 30-40 nm in diameter. This basic design feature has important implications for the microcompartmentation of ions and molecules within this organelle.

Type
Unique Approaches In Imaging, Computation and Communication for Characterization of the 3D Cell & Organelles I
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 430 - 431
Copyright
Copyright © Microscopy Society of America

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References

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7. This research is funded by NSF grant MCB-9506113 and utilizes the facilities of the Biological Microscopy and Image Reconstruction Resource which is funded by NIH grant RR01219 and NSF grant BIR-9219043.Google Scholar