Falcieri E, Mariani A R, Del Coco R, Facchini A, Maraldi N M
Istituto di Anatomia Umana Normale, Università di Bologna, Italia.
J Submicrosc Cytol Pathol. 1988 Jul;20(3):623-7.
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.
本文描述了一种用于少量分离细胞冷冻断裂的简单、高产技术。一滴悬浮固定的细胞滴在经聚赖氨酸处理的盖玻片上,通过静电力形成单层。经过冷冻保护后,将盖玻片倒置在覆盖有聚乙烯醇缩丁醛的金载体上,然后在液氮中冷冻。通过在盖玻片下推进刀片,单层细胞将被断裂。尽管细胞数量较少,但仍可暴露大面积的细胞表面,如通过流式细胞术分选细胞后可获得的细胞表面。