Electron microscopy of frozen biological suspensions

Details

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State: Public
Version: Final published version
Secondary document(s)
Sous embargo indéterminé.
State: Public
Version: Supplementary document
Serval ID
serval:BIB_4581D1BB44BD
Type
Article: article from journal or magazin.
Collection
Publications
Title
Electron microscopy of frozen biological suspensions
Journal
Journal of Microscopy
Author(s)
Lepault  J., Booy  F. P., Dubochet  J.
ISSN
0022-2720 (Print)
Publication state
Published
Issued date
01/1983
Peer-reviewed
Oui
Volume
129
Number
Pt 1
Pages
89-102
Language
english
Notes
Journal Article --- Old month value: Jan
Abstract
The methodology for preparing specimens in the frozen, hydrated state has been assessed using crystals and T4 bacteriophages. The methods have also been demonstrated with lambda bacteriophages, purple membrane of Halobacterium halobium and fibres of DNA. For particles dispersed in an aqueous environment, it is shown that optimum structural preservation is obtained from a thin, quench-frozen film with the bulk aqueous medium in the vitreous state. Crystallization of the bulk water may result in solute segregation and expulsion of the specimen from the film. Contrast measurements can be used to follow directly the state of hydration of a specimen during transition from the fully hydrated to the freeze-dried state and permit direct measurement of the water content of the specimen. By changing the concentration and composition of the aqueous medium the contrast of particles in a vitreous film can be controlled and any state of negative, positive or zero contrast may be obtained. At 100 K, frozen-hydrated, freeze-dried or sugar embedded crystals can withstand a three- to four-fold increase in electron exposure for the same damage when compared with similar sugar-embedded or freeze-dried samples at room temperature.
Keywords
Catalase Cell Membrane/ultrastructure Crystallization Dna Freeze Drying Freezing Halobacterium/ultrastructure *Microscopy, Electron *Specimen Handling Staining and Labeling T-Phages/ultrastructure Water
Pubmed
Web of science
Open Access
Yes
Create date
24/01/2008 11:25
Last modification date
20/08/2019 14:50
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