Bullivant S
J Microsc. 1977 Sep;111(1):101-16. doi: 10.1111/j.1365-2818.1977.tb00050.x.
The freeze-fracture technique is now widely used in the study of membranes, but it should be stressed that it shows internal hydrophobic planes of membranes, prepared under physical conditions far removed from those prevailing in vivo. Hence there is considerable potential for artefact. Work on the membrane lipid component, the intramembrane particles, and their aggregation under certain conditions, is reviewed in the context of fact versus artefact. Particular attention is paid to the results of complementary replica experiments, performed in the author's laboratory and elsewhere, which indicate that lipid collapse and protein particle deformation contribute to the appearance of the membrane fracture face seen in the final replica. A model, showing the effects of freeze-fracturing on membranes, is presented.
冷冻断裂技术如今在膜研究中得到广泛应用,但应当强调的是,它展示的是在与体内实际存在的物理条件相差甚远的条件下制备的膜的内部疏水平面。因此,存在相当大的人为因素影响的可能性。本文结合实际与人为因素,对膜脂质成分、膜内颗粒及其在特定条件下的聚集情况的研究进行了综述。特别关注了在作者实验室及其他地方进行的互补复制品实验的结果,这些结果表明脂质塌陷和蛋白质颗粒变形导致了最终复制品中膜断裂面的出现。本文还提出了一个展示冷冻断裂对膜影响的模型。