A site-specific focused-ion-beam lift-out method for cryo Transmission Electron Microscopy

作者：Rubino Stefano^*; Akhtar Sultan; Melin Petter; Searle Andrew; Spellward Paul; Leifer Klaus

来源：Journal of Structural Biology, 2012, 180(3): 572-576.

DOI：10.1016/j.jsb.2012.08.012

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

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.

• 出版日期2012-12