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A site-specific focused-ion-beam lift-out method for cryo Transmission Electron Microscopy
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Applied Materials Sciences.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Applied Materials Sciences.
Dept of Microbiology, Swedish University of Agricultural Sciences, Uppsala.
Gatan Inc, Abingdon Oxon, UK.
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2012 (English)In: Journal of Structural Biology, ISSN 1047-8477, E-ISSN 1095-8657, Vol. 180, no 3, 572-576 p.Article in journal (Refereed) Published
Abstract [en]

The focused-ion-beam (FIB) is the method of choice for site-specific sample preparation for Transmission Electron Microscopy (TEM) in material sciences. A lamella can be physically lifted out from a specific region of a bulk specimen with submicrometer precision and thinned to electron transparency for high-resolution imaging in the TEM. The possibility to use this tool in life sciences applications has been limited by the lack of lift-out capabilities at the cryogenic temperatures often needed for biological samples. Conventional cryo-TEM sample preparation is mostly based on ultramicrotomy, a procedure that is not site-specific and known to produce artifacts. Here we demonstrate how a cooled nanomanipulator and a custom-built transfer station can be used to achieve cryo-preparation of TEM samples with the FIB, enabling high-resolution investigation of frozen-hydrated specimens in the TEM.

Place, publisher, year, edition, pages
2012. Vol. 180, no 3, 572-576 p.
Keyword [en]
Cryo-EM, Cryo-FIB, Cryo-SEM, Cryo-TEM, FIB, Frozen-hydrated sample, Sample preparation, Specimen thinning, TEM
National Category
Natural Sciences Engineering and Technology
Research subject
Engineering Science with specialization in Materials Science
Identifiers
URN: urn:nbn:se:uu:diva-184896DOI: 10.1016/j.jsb.2012.08.012ISI: 000311471000020OAI: oai:DiVA.org:uu-184896DiVA: diva2:570911
Note

De två första författarna delar förstaförfattarskapet.

Available from: 2012-11-20 Created: 2012-11-15 Last updated: 2017-12-07Bibliographically approved

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Rubino, StefanoAkhtar, SultanLeifer, Klaus

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