FIB-SEM tomography in biology.

Détails

ID Serval
serval:BIB_5D201429E8A2
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
FIB-SEM tomography in biology.
Périodique
Methods In Molecular Biology
Auteur⸱e⸱s
Kizilyaprak C., Bittermann A.G., Daraspe J., Humbel B.M.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Statut éditorial
Publié
Date de publication
2014
Peer-reviewed
Oui
Volume
1117
Pages
541-558
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Résumé
Three-dimensional information is much easier to understand than a set of two-dimensional images. Therefore a layman is thrilled by the pseudo-3D image taken in a scanning electron microscope (SEM) while, when seeing a transmission electron micrograph, his imagination is challenged. First approaches to gain insight in the third dimension were to make serial microtome sections of a region of interest (ROI) and then building a model of the object. Serial microtome sectioning is a tedious and skill-demanding work and therefore seldom done. In the last two decades with the increase of computer power, sophisticated display options, and the development of new instruments, an SEM with a built-in microtome as well as a focused ion beam scanning electron microscope (FIB-SEM), serial sectioning, and 3D analysis has become far easier and faster.Due to the relief like topology of the microtome trimmed block face of resin-embedded tissue, the ROI can be searched in the secondary electron mode, and at the selected spot, the ROI is prepared with the ion beam for 3D analysis. For FIB-SEM tomography, a thin slice is removed with the ion beam and the newly exposed face is imaged with the electron beam, usually by recording the backscattered electrons. The process, also called "slice and view," is repeated until the desired volume is imaged.As FIB-SEM allows 3D imaging of biological fine structure at high resolution of only small volumes, it is crucial to perform slice and view at carefully selected spots. Finding the region of interest is therefore a prerequisite for meaningful imaging. Thin layer plastification of biofilms offers direct access to the original sample surface and allows the selection of an ROI for site-specific FIB-SEM tomography just by its pronounced topographic features.
Mots-clé
Electron Microscope Tomography/methods, Histocytological Preparation Techniques, Image Processing, Computer-Assisted, Microscopy, Electron, Scanning/methods, Software
Pubmed
Web of science
Création de la notice
27/08/2014 15:05
Dernière modification de la notice
20/08/2019 15:15
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