Tannic acid-mediated osmium impregnation after freeze-substitution: a strategy to enhance membrane contrast for electron tomography.
Details
Serval ID
serval:BIB_0E27F4E0D8A7
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Tannic acid-mediated osmium impregnation after freeze-substitution: a strategy to enhance membrane contrast for electron tomography.
Journal
Journal of Structural Biology
ISSN
1095-8657 (Electronic)
ISSN-L
1047-8477
Publication state
Published
Issued date
2009
Volume
166
Number
1
Pages
103-106
Language
english
Abstract
In this technical note we report a tannic acid-mediated osmium impregnation method that, applied after freeze-substitution, increases membrane contrast in cells for transmission electron microscopy and tomography studies. The general staining that is achieved allows visualization of organelles, plasma membrane and associated specializations (e.g. caveolae) in non-post-stained plastic sections by conventional transmission electron microscopy. In combination with electron tomography it results in membranes with a proper contrast and equal staining pattern through the depth of the tomograms. The protocol that we contribute can serve as starting point for those willing to improve the membrane contrast of their specimens or to make 3D studies on the architecture of membranous compartments by electron tomography.
Keywords
Buffers, Cell Line, Cell Membrane/chemistry, Cell Membrane/ultrastructure, Electron Microscope Tomography/methods, Endothelial Cells/ultrastructure, Eukaryotic Cells/ultrastructure, Fixatives/chemistry, Freeze Substitution/methods, Humans, Intracellular Membranes/chemistry, Intracellular Membranes/ultrastructure, Microscopy, Electron, Transmission, Osmium Tetroxide/chemistry, Staining and Labeling/methods, Tannins/chemistry
Pubmed
Web of science
Create date
28/02/2012 18:39
Last modification date
20/08/2019 12:35