Tannic acid-mediated osmium impregnation after freeze-substitution: a strategy to enhance membrane contrast for electron tomography.
J Struct Biol
; 166(1): 103-6, 2009 Apr.
Article
em En
| MEDLINE
| ID: mdl-19162195
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Tetróxido de Ósmio
/
Coloração e Rotulagem
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Taninos
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Membrana Celular
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Substituição ao Congelamento
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Tomografia com Microscopia Eletrônica
Limite:
Humans
Idioma:
En
Ano de publicação:
2009
Tipo de documento:
Article