Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy.

Details

Serval ID
serval:BIB_5750EB83FE03
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy.
Journal
Journal of visualized experiments
Author(s)
Franke T., Kolotuev I.
ISSN
1940-087X (Electronic)
ISSN-L
1940-087X
Publication state
Published
Issued date
15/07/2021
Peer-reviewed
Oui
Number
173
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't ; Video-Audio Media
Publication Status: epublish
Abstract
Electron microscopy is applied in biology and medicine for imaging of cellular and structural details at nanometer resolution. Historically, Transmission Electron Microscopy (TEM) provided insight into cell ultrastructure, but in the recent decade, the development of modern Scanning Electron Microscopes (SEM) has changed the way of looking inside the cells. Even though the resolution of TEM is superior when protein-level structural details are needed, SEM-resolution is sufficient for the majority of the organelle-level cell biology-related questions. The advancement in technology enabled automatic volume acquisition solutions such as Serial block-face imaging (SBF-SEM) and Focused ion beam SEM (FIB-SEM). Nevertheless, to this day, these methods remain inefficient when the identification and navigation to areas of interest are crucial. Without the means for precise localization of target areas before imaging, operators need to acquire much more data than they need (in SBF-SEM), or, even worse, prepare many grids and image them all (in TEM). We propose the strategy of "lateral screening" using Array Tomography in SEM, which facilitates the localization of areas of interest, followed by automated imaging of the relevant fraction of the total sample volume. Array tomography samples are conserved during imaging, and they can be arranged into section libraries ready for repeated imaging. Several examples are shown in which lateral screening enables us to analyze structural details that are incredibly challenging to access with any other method.
Keywords
Imaging, Three-Dimensional, Microscopy, Electron, Scanning, Microscopy, Electron, Transmission, Tomography, Workflow
Pubmed
Open Access
Yes
Create date
01/10/2021 9:39
Last modification date
29/07/2022 6:38
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