DOI: 10.1111/j.1365-2818.1991.tb03087.x. Preparation of ultrathin amorphous ice films for cryo‐electron microscopy

Preparation of ultrathin amorphous ice films for cryo‐electron microscopy

10.1111/j.1365-2818.1991.tb03087.x

Crossref journal-article

Wiley

Journal of Microscopy (311)

Abstract

SUMMARYElectron microscopy of biological macromolecules embedded in vitrified ice films suffers from serious problems caused by excessive inelastic scattering, beam‐induced movements of the specimen, deformation of the molecules by adsorption at the water‐air interface and insufficient mechanical stability of the films. We have built an environmental chamber to control temperature and humidity independently in order to produce ultrathin water films (< 20 nm) spanning holes with diameters of 20 μm to 1 mm. The surface tension of the water films was reduced by adding lipid monolayers, thus prolonging the usable time for thinning of the film and avoiding adsorption artefacts in the embedded material. After cryofixation in ethane a carbon film was evaporated on each side of the specimen to stabilize the ultrathin ice—lipid film. Mechanical stability and charging effects could successfully be reduced by this preparation method. Collapsing water films could be cryofixed and the shape of the hole was analysed. By the eccentricity of the elliptical holes an estimation could be made of the burst velocity of the rim of the hole and of the cooling rate of the cryofixation process.

Bibliography

Jakubowski, U., & Mende, M. (1991). Preparation of ultrathin amorphous ice films for cryoâelectron microscopy. Journal of Microscopy, 161(2), 241â252. Portico.

Authors 2

U. Jakubowski (first)
M. Mende (additional)

References 11 Referenced 9

10.1002/jemt.1060100111
10.1016/0968-0004(85)90150-1
10.1111/j.1365-2818.1982.tb04626.x
10.1007/978-1-4684-2958-9
{'issue': '3', 'key': 'e_1_2_1_6_1', 'first-page': '277', 'article-title': 'Thin film formation and the imaging of phospholipid by cryo‐electron microscopy', 'author': 'Frederik P.M.', 'year': '1989', 'journal-title': 'Scanning Microsc.'} / Scanning Microsc. / Thin film formation and the imaging of phospholipid by cryo‐electron microscopy by Frederik P.M. (1989)
10.1016/0304-3991(89)90333-1
10.1007/978-3-642-72815-0_2
10.1021/j100843a042
10.1016/0304-3991(85)90102-0
10.1002/jemt.1060050308
10.1111/j.1365-2818.1986.tb02714.x

Dates

Type	When
Created	14 years ago (Aug. 2, 2011, 1:17 p.m.)
Deposited	1 year, 10 months ago (Oct. 23, 2023, 3:14 a.m.)
Indexed	1 year, 10 months ago (Oct. 29, 2023, 4:20 a.m.)
Issued	34 years, 6 months ago (Feb. 1, 1991)
Published	34 years, 6 months ago (Feb. 1, 1991)
Published Online	14 years ago (Aug. 2, 2011)
Published Print	34 years, 6 months ago (Feb. 1, 1991)

Funders 0

None

BibTeX

@article{Jakubowski_1991, title={Preparation of ultrathin amorphous ice films for cryo‐electron microscopy}, volume={161}, ISSN={1365-2818}, url={http://dx.doi.org/10.1111/j.1365-2818.1991.tb03087.x}, DOI={10.1111/j.1365-2818.1991.tb03087.x}, number={2}, journal={Journal of Microscopy}, publisher={Wiley}, author={Jakubowski, U. and Mende, M.}, year={1991}, month=feb, pages={241–252} }

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