title: The sleeping beauty kissed awake: new methods in electron microscopy to study cellular membranes.
abstract: Electron microscopy (EM) for biological samples, developed in the 1940-1950s, changed our conception about the architecture of eukaryotic cells. It was followed by a period where EM applied to cell biology had seemingly fallen asleep, even though new methods with important implications for modern EM were developed. Among these was the discovery that samples can be preserved by chemical fixation and most importantly by rapid freezing without the formation of crystalline ice, giving birth to the world of cryo-EM. The past 15-20 years are hallmarked by a tremendous interest in EM, driven by important technological advances. Cryo-EM, in particular, is now capable of revealing structures of proteins at a near-atomic resolution owing to improved sample preparation methods, microscopes and cameras. In this review, we focus on the challenges associated with the imaging of membranes by EM and give examples from the field of host-pathogen interactions, in particular of virus-infected cells. Despite the advantages of imaging membranes under native conditions in cryo-EM, conventional EM will remain an important complementary method, in particular if large volumes need to be imaged.
authors.affiliation: Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892, U.S.A. petr.chlanda@nih.gov jacomina.krijnse-locker@pasteur.fr.
authors.forename: Petr
authors.initials: P
authors.lastname: Chlanda
authors.affiliation: Ultrapole, Ultra-Structural Bio-Imaging, Institut Pasteur, 28, rue du Dr. Roux, Paris 75015, France petr.chlanda@nih.gov jacomina.krijnse-locker@pasteur.fr.
authors.forename: Jacomine
authors.initials: J
authors.lastname: Krijnse Locker
mesh_terms: D000180:Acrylic Resins; D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure* / Q000821:virology; D002696:Chlamydomonas / Q000648:ultrastructure*; D020285:Cryoelectron Microscopy / Q000266:history / Q000295:instrumentation / Q000379:methods*; D006367:HeLa Cells; D049673:History, 20th Century; D049674:History, 21st Century; D054884:Host-Pathogen Interactions; D006801:Humans; D021621:Imaging, Three-Dimensional; D051379:Mice; D009900:Optic Nerve / Q000648:ultrastructure*; D014616:Vaccinia virus / Q000648:ultrastructure*; D014771:Virion / Q000648:ultrastructure*; D058989:Vitrification
chemical_list: D000180:Acrylic Resins; C037883:Lowicryl 4KM resin
keywords: electron microscopy; enveloped viruses; membranes