title: Making the practically impossible "Merely difficult"--Cryogenic FIB lift-out for "Damage free" soft matter imaging.
abstract: The preparation of thinned lamellae from bulk samples for transmission electron microscopy (TEM) analysis has been possible in the focussed ion beam scanning electron microscope (FIB-SEM) for over 20 years via the in situ lift-out method. Lift-out offers a fast and site specific preparation method for TEM analysis, typically in the field of materials science. More recently it has been applied to a low-water content biological sample (Rubino 2012). This work presents the successful lift-out of high-water content lamellae, under cryogenic conditions (cryo-FIB lift-out) and using a nanomanipulator retaining its full range of motion, which are advances on the work previously done by Rubino (2012). Strategies are explored for maintaining cryogenic conditions, grid attachment using cryo-condensation of water and protection of the lamella when transferring to the TEM.
authors.affiliation: Nottingham Nanoscale and Microscale Research Centre, University of Nottingham, United Kingdom.
authors.forename: Christopher D J
authors.initials: CD
authors.lastname: Parmenter
authors.affiliation: Nottingham Nanoscale and Microscale Research Centre, University of Nottingham, United Kingdom.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Fay
authors.affiliation: Oxford Instruments Nanoanalysis, Concord, MA.
authors.forename: Cheryl
authors.initials: C
authors.lastname: Hartfield
authors.affiliation: Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), School of Pharmacy, University of Nottingham, United Kingdom.
authors.forename: Hoda M
authors.initials: HM
authors.lastname: Eltaher
mesh_terms: D016616:Cryoultramicrotomy / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation*; D014867:Water / Q000737:chemistry
chemical_list: D014867:Water
keywords: biological-EM; cryo-FIB; cryo-TEM preparation; cryo-lift-out; frozen-hydrated; soft-matter