Serial block face-scanning electron microscopy for volume electron microscopy

Saskia Lippens; Anna Kremer; Peter Borghgraef; Christopher J Guérin

doi:10.1016/bs.mcb.2019.04.002

Serial block face-scanning electron microscopy for volume electron microscopy

Methods Cell Biol. 2019:152:69-85. doi: 10.1016/bs.mcb.2019.04.002. Epub 2019 Jun 17.

Authors

Saskia Lippens¹, Anna Kremer², Peter Borghgraef², Christopher J Guérin²

Affiliations

¹ VIB BioImaging Core, VIB, Ghent, Belgium; VIB Inflammation Research Center, VIB, Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium. Electronic address: saskia.lippens@irc.vib-ugent.be.
² VIB BioImaging Core, VIB, Ghent, Belgium; VIB Inflammation Research Center, VIB, Ghent, Belgium; Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.

PMID: 31326027
DOI: 10.1016/bs.mcb.2019.04.002

Abstract

There are different technologies that can be used to obtain a 3D image at nanometer resolution. Over the past decade, there has been a growing interest in applying Serial Block Face Scanning Electron Microscopy (SBF-SEM) in different fields of life science research. This technology has the advantage that it can cover a range of volumes, going from monolayers to multiple tissue layers in all three dimensions. SBF-SEM was originally used in neuroscience and then expanded to other research domains. The whole process of sample preparation for SBF-SEM is very long and consists of many steps, which makes adjustment of a given workflow very challenging. Here we describe the SBF-SEM workflow and those steps in the process that can be tweaked for any sample.

Keywords: Focal charge compensator; Sample preparation; Serial block-face scanning electron microscopy; Three dimensional electron microscopy; Volume electron microscopy.

MeSH terms

Imaging, Three-Dimensional / methods
Microscopy, Electron, Scanning / methods*