title: A site-specific focused-ion-beam lift-out method for cryo Transmission Electron Microscopy.
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.
authors.affiliation: Department of Engineering Sciences, Uppsala University, Box 534, 751 21 Uppsala, Sweden. stefanorubino@yahoo.it
authors.forename: Stefano
authors.initials: S
authors.lastname: Rubino
authors.forename: Sultan
authors.initials: S
authors.lastname: Akhtar
authors.forename: Petter
authors.initials: P
authors.lastname: Melin
authors.forename: Andrew
authors.initials: A
authors.lastname: Searle
authors.forename: Paul
authors.initials: P
authors.lastname: Spellward
authors.forename: Klaus
authors.initials: K
authors.lastname: Leifer
mesh_terms: D002118:Calcium; D002413:Cations, Divalent; D020285:Cryoelectron Microscopy / Q000295:instrumentation* / Q000379:methods; D011211:Electric Power Supplies; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation* / Q000379:methods; D013048:Specimen Handling / Q000295:instrumentation* / Q000379:methods
chemical_list: D002413:Cations, Divalent; D002118:Calcium