A simplified 'sandwich' technique for in situ embedding and perpendicular sectioning of monolayer cultures of human skin fibroblasts.
In the processing of cell cultures, grown as a monolayer in tissue culture dishes for electron microscopy, the sectioning of the monolayer is an essential step. The monolayer can be sectioned either parallel or perpendicular to the plane of growth. Several methods for the perpendicular way of sectioning have already been described. We propose a simplified method in which the monolayer is sandwiched between two layers of resin, one of which is a prepolymerized block, the other being a layer of resin, applied at a second stage. Sectioning of this 'flat embedded' specimen yields thin sections perpendicular to the plane of growth of the monolayer without elaborate orientating procedures. The advantage of this procedure is that it can be done using only routine embedding techniques, avoiding special materials or complex manipulations. This sandwich technique provides an excellent mechanical fixation of the monolayer and protects it against external damage.
Govaerts
P J
PJ
University of Antwerp (UIA), Department of Pediatrics, Antwerp-Wilrijk, Belgium.
Bernaert
I I
II
du Caju
M V
MV
Jacob
W A
WA
D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods; D005347:Fibroblasts / Q000166:cytology / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron / Q000379:methods; D012867:Skin / Q000166:cytology* / Q000648:ultrastructure