Research article - Peer-reviewed, 2012
A site-specific focused-ion-beam lift-out method for cryo Transmission Electron Microscopy
Rubino, Stefano; Akhtar, Sultan; Melin, Petter; Searle, Andrew; Spellward, Paul; Leifer, KlausAbstract
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. (C) 2012 Elsevier Inc. All rights reserved.Keywords
Cryo-SEM; Cryo-FIB; Cryo-TEM; Specimen thinning; Frozen-hydrated sample; Cryo-EM; FIB; TEM; Sample preparationPublished in
Journal of Structural Biology2012, volume: 180, number: 3, pages: 572-576
Publisher: ACADEMIC PRESS INC ELSEVIER SCIENCE
Authors' information
Rubino, Stefano
Akhtar, Sultan
Melin, Petter
Swedish University of Agricultural Sciences, Department of Microbiology
Searle, Andrew
Spellward, Paul
Leifer, Klaus
UKÄ Subject classification
Microbiology
Publication Identifiers
DOI: https://doi.org/10.1016/j.jsb.2012.08.012
URI (permanent link to this page)
https://res.slu.se/id/publ/42408