Vitrification of cryoelectron microscopy specimens revealed by high-speed photographic imaging.
Details
Serval ID
serval:BIB_28888
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Vitrification of cryoelectron microscopy specimens revealed by high-speed photographic imaging.
Journal
Journal of Microscopy
ISSN
0022-2720
Publication state
Published
Issued date
2003
Peer-reviewed
Oui
Volume
211
Number
Pt 1
Pages
48-53
Language
english
Abstract
Cryoelectron microsopy is a widely used technique to observe biological material in an almost physiological, fully hydrated state. The sample is prepared for electron microsopy observation by quickly reducing its temperature to -180 degrees C. The high-speed cooling induces the formation of vitreous water, which preserves the sample conformation. However, the way vitrification occurs is still poorly understood. In order to better understand the phenomenon, we have used a stroboscopic device to visualize the interaction between the electron microscopy grid and the cryogen. By blocking the free fall of the plunger once the grid has penetrated the coolant by half its diameter, we have elucidated the way in which vitrification propagates. The findings were confirmed by numerical simulation. In addition, according to our observations, we now present an alternative way to prepare vitreous specimens. This new method, with the grid parallel to the liquid cryogen surface, decreases evaporation from the sample during its free fall towards the coolant and at the same time achieves a more uniform vitrification over the entire surface of the specimen.
Keywords
Cryoelectron Microscopy, Freezing, Image Processing, Computer-Assisted, Photography/instrumentation, Specimen Handling, Time Factors
OAI-PMH
Pubmed
Web of science
Create date
19/11/2007 12:26
Last modification date
20/08/2019 13:08