title: Volume scanning electron microscopy for imaging biological ultrastructure.
abstract: Electron microscopy (EM) has been a key imaging method to investigate biological ultrastructure for over six decades. In recent years, novel volume EM techniques have significantly advanced nanometre-scale imaging of cells and tissues in three dimensions. Previously, this had depended on the slow and error-prone manual tasks of cutting and handling large numbers of sections, and imaging them one-by-one with transmission EM. Now, automated volume imaging methods mostly based on scanning EM (SEM) allow faster and more reliable acquisition of serial images through tissue volumes and achieve higher z-resolution. Various software tools have been developed to manipulate the acquired image stacks and facilitate quantitative analysis. Here, we introduce three volume SEM methods: serial block-face electron microscopy (SBEM), focused ion beam SEM (FIB-SEM) and automated tape-collecting ultramicrotome SEM (ATUM-SEM). We discuss and compare their capabilities, provide an overview of the full volume SEM workflow for obtaining 3D datasets and showcase different applications for biological research.
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland. benjamin.titze@fmi.ch.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Titze
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
authors.forename: Christel
authors.initials: C
authors.lastname: Genoud
mesh_terms: D000818:Animals; D001921:Brain / Q000648:ultrastructure*; D016591:Histocytological Preparation Techniques / Q000379:methods; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D008867:Microtomy / Q000379:methods
keywords: Brain/nervous system; Cellular imaging; Electron microscopy; Systems biology