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Use of cryo-negative staining in tomographic reconstruction of biological objects: application to T4 bacteriophage.
- 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.
- Subjects :
- Bacteriophage T4 isolation & purification
Cryoelectron Microscopy methods
Escherichia coli pathogenicity
Image Processing, Computer-Assisted
Imaging, Three-Dimensional
Specimen Handling
Bacteriophage T4 ultrastructure
Cryoelectron Microscopy instrumentation
Negative Staining methods
Tomography
Subjects
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