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Research article2012Peer reviewed

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, Klaus


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.


Cryo-SEM; Cryo-FIB; Cryo-TEM; Specimen thinning; Frozen-hydrated sample; Cryo-EM; FIB; TEM; Sample preparation

Published in

Journal of Structural Biology
2012, Volume: 180, number: 3, pages: 572-576

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