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          Institute: MPI für Biochemie     Collection: Structural Biology (W. Baumeister)     Display Documents

ID: 318322.0, MPI für Biochemie / Structural Biology (W. Baumeister)
Electron tomography of ice-embedded prokaryotic cells
Authors:Grimm, R.; Singh, H.; Rachel, R.; Typke, D.; Zillig, W.; Baumeister, W.
Date of Publication (YYYY-MM-DD):1998-02
Title of Journal:Biophysical Journal
Issue / Number:2 Part 1
Start Page:1031
End Page:1042
Audience:Not Specified
Abstract / Description:Whole cells of archaea were embedded in vitreous ice by plunge freezing and investigated by automated energy-filtered electron tomography at 120 kV. The embedded cells were between 300 and 750 nm thick, and their structures were reconstructed to a resolution of 20-40 nm from tilt series comprising 50-140 images, The dose was kept within tolerable limits. A resolution of 20 nm allowed visualization of the individual stalks of the S-layer of Pyrobaculum aerophilum cells, which had undergone partial lysis, in three dimensions. The attainable resolution for low-dose electron tomography under different experimental conditions was theoretically investigated in terms of the specimen thickness. To obtain 2-nm resolution at 120 kV (300 kV), the specimen must not be thicker than 100 nm (150 nm). For a resolution of 10 nm, the maximum thickness is 450 nm (700 nm). An accelerating voltage of 300 kV is advantageous, mainly for specimens thicker than 100 nm. Experimental investigations so far have resulted in a resolution that is worse by a factor of 2-5 as compared to theory. [References: 37]
Free Keywords:3-dimensional structure; Biological applications; Microscopy; Reconstruction; Specimens; Sulfolobus; Vesicles; Envelope; Filter; Layer.; Biochemistry & biophysics.
External Publication Status:published
Document Type:Article
Communicated by:Anton Hillebrand
Affiliations:MPI für Biochemie/Structural Biology (W. Baumeister)/
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