Abstract

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 preparation

Published in

Journal of Structural Biology
2012, volume: 180, number: 3, pages: 572-576
Publisher: ACADEMIC PRESS INC ELSEVIER SCIENCE

Authors' information