Scanning transmission electron microscopy (STEM) allows atomic scale characterization of solid-solid interfaces, but has seen limited applications to solid-liquid interfaces due to the volatility of liquids in the microscope vacuum. Although cryo-electron microscopy is routinely used to characterize hydrated samples stabilized by rapid freezing, sample thinning is required to access the internal interfaces of thicker specimens. Here, we adapt cryo-focused ion beam (FIB) "lift-out," a technique recently developed for biological specimens, to prepare intact internal solid-liquid interfaces for high-resolution structural and chemical analysis by cryo-STEM. To guide the milling process we introduce a label-free in situ method of localizing subsurface structures in suitable materials by energy dispersive X-ray spectroscopy (EDX). Monte Carlo simulations are performed to evaluate the depth-probing capability of the technique, and show good qualitative agreement with experiment. We also detail procedures to produce homogeneously thin lamellae, which enable nanoscale structural, elemental, and chemical analysis of intact solid-liquid interfaces by analytical cryo-STEM. This work demonstrates the potential of cryo-FIB lift-out and cryo-STEM for understanding physical and chemical processes at solid-liquid interfaces.
Keywords: cryo-focused ion beam lift-out; cryo-scanning transmission electron microscopy; solid–liquid interfaces; spatially resolved electron energy loss spectroscopy.
Cryo-STEM Mapping of Solid-Liquid Interfaces and Dendrites in Lithium-Metal BatteriesMJ Zachman et al. Nature 560 (7718), 345-349. PMID 30111789.Solid-liquid interfaces are important in a range of chemical, physical and biological processes1-4, but are often not fully understood owing to the lack of hig …
A Focused Ion Beam Milling and Lift-Out Approach for Site-Specific Preparation of Frozen-Hydrated Lamellas From Multicellular OrganismsJ Mahamid et al. J Struct Biol 192 (2), 262-9. PMID 26216184.Cryo-electron tomography provides 3D views of cellular architecture with molecular resolution. A principal limitation of cryo-transmission electron microscopy performed o …
Cryo-focused-ion-beam Applications in Structural BiologyA Rigort et al. Arch Biochem Biophys 581, 122-30. PMID 25703192. - ReviewThe ability to precisely control the preparation of biological samples for investigations by electron cryo-microscopy is becoming increasingly important for ultrastructur …
Making the Practically Impossible "Merely difficult"--Cryogenic FIB Lift-Out for "Damage Free" Soft Matter ImagingCD Parmenter et al. Microsc Res Tech 79 (4), 298-303. PMID 26875988.The preparation of thinned lamellae from bulk samples for transmission electron microscopy (TEM) analysis has been possible in the focussed ion beam scanning electron mic …
Opening Windows Into the Cell: Focused-Ion-Beam Milling for Cryo-Electron TomographyE Villa et al. Curr Opin Struct Biol 23 (5), 771-7. PMID 24090931. - ReviewCryo-electron tomography (CET) is ideally suited for bridging the resolution gap between molecular and cellular structural studies. However, CET is limited to a sample th …
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In Situ Microfluidic Cryofixation for Cryo Focused Ion Beam Milling and Cryo Electron TomographyM Fuest et al. Sci Rep 9 (1), 19133. PMID 31836773.We present a microfluidic platform for studying structure-function relationships at the cellular level by connecting video rate live cell imaging with in situ microfluidi …