Quick-freeze, deep-etch replication of cells in monolayers.
J Electron Microsc Tech
; 14(4): 342-7, 1990 Apr.
Article
en En
| MEDLINE
| ID: mdl-2332810
We have made several technical improvements for quick-freeze, deep-etch replication of monolayers of cells grown on, or attached to, glass coverslips. Cells studied include muscle cells of rat and Xenopus cultured on glass coverslips, and erythrocytes attached to coverslips coated with poly-L-lysine. We describe methods for identifying particular areas of cultures, e.g., clusters of acetylcholine receptors on muscle cells, by light microscopy and then relocating these areas after replication. For good preservation of structure by quick-freezing, it is necessary to ensure that the surface to be frozen is covered by a minimum depth of water (less than 10 microns). Insufficient or excess water left on the sample during freezing causes recognizable artifacts in its replica. We describe two ways to control the water table--one by improving visual control of water removal, the other by blowing excess water off the sample surface with a jet of nitrogen applied during its descent to the freezing block. Finally, we describe a new specimen holder that allows us to etch and replicate six samples at once with good thermal contact between the stage and samples.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Técnica de Fractura por Congelación
/
Músculos
Límite:
Animals
Idioma:
En
Revista:
J Electron Microsc Tech
Asunto de la revista:
DIAGNOSTICO POR IMAGEM
Año:
1990
Tipo del documento:
Article
Pais de publicación:
Estados Unidos