Volume scanning electron microscopy for imaging biological ultrastructure.

Titze B; Genoud C

doi:10.1111/boc.201600024

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Journal article

Volume scanning electron microscopy for imaging biological ultrastructure.

Titze B Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland. benjamin.titze@fmi.ch.
Genoud C Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.

2016-07-20

Published in:

Biology of the cell. - 2016

Histocytological Preparation Techniques

Image Processing, Computer-Assisted

Microscopy, Electron, Scanning

English 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.

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