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Use of cryo-negative staining in tomographic reconstruction of biological objects: application to T4 bacteriophage.

Authors :
Messaoudi C
Boudier T
Lechaire JP
Rigaud JL
Delacroix H
Gaill F
Marco S
Source :
Biology of the cell [Biol Cell] 2003 Sep; Vol. 95 (6), pp. 393-8.
Publication Year :
2003

Abstract

Recent advances in electron microscopy and image analysis techniques have resulted in the development of tomography, which makes possible the study of structures neither accessible to X-ray crystallography nor nuclear magnetic resonance. However, the use of tomography to study biological structures, ranging from 100 to 500 nm, requires developments in sample preparation and image analysis. Indeed, cryo-electron tomography present two major drawbacks: the low contrast of recorded images and the sample radiation damage. In the present work we have tested, on T4 bacteriophage samples, the use of a new preparation technique, cryo-negative staining, which reduces the radiation damage while preserving a high signal-to-noise ratio. Our results demonstrate that the combination of cryo-negative staining in tomography with standard cryo-microscopy and single particle analysis results in a methodological approach that could be useful in the study of biological structures ranging in the T4 bacteriophage size.

Details

Language :
English
ISSN :
0248-4900
Volume :
95
Issue :
6
Database :
MEDLINE
Journal :
Biology of the cell
Publication Type :
Academic Journal
Accession number :
14519556
Full Text :
https://doi.org/10.1016/s0248-4900(03)00086-8