Aclar discs: a versatile substrate for routine high-pressure freezing of mammalian cell monolayers.

Détails

ID Serval
serval:BIB_8F65FE21CAD3
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Aclar discs: a versatile substrate for routine high-pressure freezing of mammalian cell monolayers.
Périodique
Journal of Microscopy
Auteur(s)
Jiménez N., Humbel B.M., van Donselaar E., Verkleij A.J., Burger K.N.
ISSN
0022-2720 (Print)
ISSN-L
0022-2720
Statut éditorial
Publié
Date de publication
2006
Volume
221
Numéro
Pt 3
Pages
216-223
Langue
anglais
Résumé
High-pressure freezing avoids the artefacts induced by conventional chemical fixation, and, in combination with freeze-substitution and plastic embedding, is a reliable method for the ultrastructural analysis of mammalian cell monolayers. In order to high-pressure freeze mammalian cell monolayers, cells have to be seeded on a suitable substrate. Unfortunately, electron microscopy analysis is often hampered by poor cell growth, changes in cell morphology induced by the cell substrate or cell loss during processing. We report a method to culture, high-pressure freeze, freeze-substitute and plastic embed mammalian cell monolayers. The method is based on the use of Aclar, a copolymer film with properties very similar to those of tissue culture plastic. We show that Aclar discs support the normal growth and morphology of a wide variety of mammalian cell types, and form an ideal starting point for high-pressure freezing, freeze-substitution and plastic embedding. We present a complete protocol, which, because of its simplicity and reproducibility, provides a method suitable for the routine analysis of mammalian cell monolayers by electron microscopy and tomography.
Mots-clé
Animals, Caco-2 Cells, Cryopreservation/instrumentation, Cryopreservation/methods, Freeze Substitution, HeLa Cells, Humans, Pressure
Pubmed
Web of science
Création de la notice
28/02/2012 19:51
Dernière modification de la notice
03/03/2018 19:21
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