Pinto da Silva P, Parkison C, Dwyer N
Proc Natl Acad Sci U S A. 1981 Jan;78(1):343-7. doi: 10.1073/pnas.78.1.343.
A method--"fracture label"--is described for the cytochemical labeling of the membrane faces produced by freeze-fracture. Human erythrocytes embedded in a crosslinked matrix are frozen, fractured in liquid nitrogen, thawed, labeled, and cut into thin sections. Electron microscope observation of the fracture faces shows preferential partition of concanavalin A binding sites with the inner half of the membrane. This signifies that, during freeze-fracture, binding sites are dragged from the outer surface across the outer ("exoplasmic") half of the membrane and retained on the protoplasmic fracture face (face P). The fracture process results in exposure of new anionic sites on face P. Fracture-label can be applied to the cytochemical characterization of the cellular components exposed by freeze-fracture of isolated cells and tissues.
本文描述了一种用于对冷冻断裂产生的膜面进行细胞化学标记的方法——“断裂标记”。将包埋在交联基质中的人红细胞冷冻，在液氮中进行断裂，解冻，标记，然后切成薄片。对断裂面的电子显微镜观察显示，伴刀豆球蛋白A结合位点优先分布在膜的内半部分。这表明，在冷冻断裂过程中，结合位点从外表面被拖过膜的外（“胞质外”）半部分，并保留在原生质断裂面（P面）上。断裂过程导致P面上新的阴离子位点暴露。断裂标记可用于对分离的细胞和组织冷冻断裂后暴露的细胞成分进行细胞化学表征。
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