Tsuji Takuma, Fujita Akikazu, Fujimoto Toyoshi
Department of Anatomy and Molecular Cell Biology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Field of Veterinary Pathobiology, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan.
Methods Mol Biol. 2018;1804:231-239. doi: 10.1007/978-1-4939-8552-4_11.
Because chemical fixatives like aldehydes do not work on most lipid molecules in the membrane, small-scale lipid distribution cannot be identified by immunoelectron microscopy in cells fixed by conventional methods. Here we describe a method for physically stabilizing membranes through quick-freezing and freeze-fracture replica formation and for specifically labeling gangliosides for electron microscopy. This method enables the ultrahigh-resolution mapping of membrane lipids including gangliosides within the two-dimensional plane of membranes.
由于醛类等化学固定剂对膜中的大多数脂质分子不起作用，因此在通过传统方法固定的细胞中，免疫电子显微镜无法识别小规模的脂质分布。在此，我们描述了一种通过快速冷冻和冷冻断裂复制品形成来物理稳定膜，并对神经节苷脂进行特异性标记以用于电子显微镜观察的方法。该方法能够在膜的二维平面内对包括神经节苷脂在内的膜脂质进行超高分辨率定位。
