title: Imaging protein structure in water at 2.7 nm resolution by transmission electron microscopy.
abstract: We demonstrate an in situ transmission electron microscopy technique for imaging proteins in liquid water at room temperature. Liquid samples are loaded into a microfabricated environmental cell that isolates the sample from the vacuum with thin silicon nitride windows. We show that electron micrographs of acrosomal bundles in water are similar to bundles imaged in ice, and we determined the resolution to be at least 2.7 nm at doses of ∼35 e/Å(2). The resolution was limited by the thickness of the window and radiation damage. Surprisingly, we observed a smaller fall-off in the intensity of reflections in room-temperature water than in 98 K ice. Thus, our technique extends imaging of unstained and unlabeled macromolecular assemblies in water from the resolution of the light microscope to the nanometer resolution of the electron microscope. Our results suggest that real-time imaging of protein dynamics is conceptually feasible.
authors.affiliation: Mechanobiology Institute, National University of Singapore, Singapore.
authors.forename: Utkur M
authors.initials: UM
authors.lastname: Mirsaidov
authors.forename: Haimei
authors.initials: H
authors.lastname: Zheng
authors.forename: Yosune
authors.initials: Y
authors.lastname: Casana
authors.forename: Paul
authors.initials: P
authors.lastname: Matsudaira
mesh_terms: D000199:Actins / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D012825:Silicon / Q000737:chemistry; D014867:Water / Q000737:chemistry*
chemical_list: D000199:Actins; D014867:Water; D012825:Silicon