A high yield technique for freeze-fracturing of small fractions of isolated cells.
A simple, high-yield technique for the freeze-fracturing of small amounts of isolated cells is described. A drop of cells fixed in suspension is deposited on a polylysine-treated coverslip, forming a monolayer through electrostatic forces. After cryoprotection, the coverslip is inverted on a gold carrier covered with Vinol and then frozen in liquid nitrogen. The monolayer will be fractured by advancing the knife under the coverslip. Large areas of cell surface can be exposed despite their low number, such as that obtainable after cell sorting by flow cytometry.
Falcieri
E
E
Istituto di Anatomia Umana Normale, Università di Bologna, Italia.
Mariani
A R
AR
Del Coco
R
R
Facchini
A
A
Maraldi
N M
NM
D004912:Erythrocytes / Q000648:ultrastructure; D005347:Fibroblasts / Q000648:ultrastructure; D005614:Freeze Fracturing / Q000379:methods*; D006801:Humans; D008214:Lymphocytes / Q000648:ultrastructure; D008854:Microscopy, Electron