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Preparation of holey carbon films suitable for cryo-electron microscopy

✍ Scribed by Murray, John M. ;Ward, Rob

Publisher
Wiley (John Wiley & Sons)
Year
1987
Tongue
English
Weight
541 KB
Volume
5
Category
Article
ISSN
0741-0581

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✦ Synopsis

We present a method for the quick and reliable production of carbon films perforated with 5-10-pm holes, suitable for supporting suspensions that are to be rapidly frozen and imaged, using cryo-electron microscopy, in the frozen-hydrated state. The holes support biological specimens that are often embedded in 50-200 nm of vitreous ice, which is optimal for image contrast and preservation of large macromolecular complexes.

Holey support films, Frozen-hydrated suspensions

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Evaporation of water cannot be fully avoided when a n unsupported thin vitrified film of a n aqueous suspension is prepared for cryo-electron microscopy. This results in increasing concentration of solute which could affect the observed material. We have quantitatively studied this effect by measuri