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Vitrification of cryoelectron microscopy specimens revealed by high‐speed photographic imaging
Author(s) -
Kasas S.,
Dumas G.,
Dietler G.,
Catsicas S.,
Adrian M.
Publication year - 2003
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1046/j.1365-2818.2003.01193.x
Subject(s) - vitrification , coolant , plunger , materials science , evaporation , microscopy , electron microscope , sample (material) , scanning electron microscope , sample preparation , analytical chemistry (journal) , optics , composite material , chemistry , chromatography , thermodynamics , physics , nuclear physics
Summary 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 °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.