On the preparation of cryosections for immunocytochemistry

Détails

ID Serval
serval:BIB_50AB7E3DD0D5
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
On the preparation of cryosections for immunocytochemistry
Périodique
Journal of Ultrastructure Research
Auteur(s)
Griffiths  G., McDowall  A., Back  R., Dubochet  J.
ISSN
0022-5320 (Print)
Statut éditorial
Publié
Date de publication
10/1984
Volume
89
Numéro
1
Pages
65-78
Notes
Journal Article --- Old month value: Oct
Résumé
The key preparation steps in the Tokuyasu thawed frozen section technique for immunocytochemistry, namely freezing, sectioning, thawing, and drying, were studied. A spherical tissue culture cell was used as a model system. The frozen hydrated section technique indicated that glutaraldehyde-fixed, 2.1 M sucrose-infused pellets of cells were routinely vitrified by immersion in liquid nitrogen but water was crystallized when lower sucrose concentrations (0.6-1 M) were used. Quantitative mass measurements showed that the fixed cells are freely permeable to sucrose. The frozen hydrated sections were severely compressed but cell profiles regained their circular appearance upon thawing. The average section thickness of our frozen-hydrated sections was 110 nm: this was reduced to 30-50 nm upon thawing, washing, and air-drying. This change was accompanied by severe drying artifacts. By using the methyl cellulose drying technique, this collapse upon air-drying could be significantly reduced, but not completely prevented, giving an average thickness of 70 nm.
Mots-clé
Animals Cell Line Cricetinae Cricetulus Female *Frozen Sections Immunochemistry/*methods Microscopy, Electron *Microtomy Ovary
Pubmed
Web of science
Création de la notice
24/01/2008 11:25
Dernière modification de la notice
03/03/2018 17:09
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