Okanoue T, Ohta M, Kachi K, Ohta Y, Kanaoka H, Sawa Y, Kagawa K, Takino T, French S W
Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Japan.
J Hepatol. 1988 Jun;6(3):291-8. doi: 10.1016/s0168-8278(88)80045-x.
Portions of eight human livers taken by wedge biopsy or needle biopsy were extracted with 0.5% Triton X-100 and then studied by scanning electron microscopy and immunoelectron microscopy. The wedge-biopsy specimens were perfused with the detergent solution. Needle-biopsy specimens were quickly frozen and cracked and then the cracked tissue was immersed in the detergent solution. The three-dimensional filament network of hepatocytes was visualized. A dense network which consisted of intermediate filaments and microfilaments was observed within the cytoplasm of hepatocytes. These filaments were better preserved in the needle biopsies which were quick-frozen and cracked before extraction than in the wedge biopsies. Variation in the amount of the cytoskeletal filaments was less prominent in the hepatocytes treated by rapid freezing. It is concluded that freeze-cracking is the most favorable method for the study of cytoskeletal pathology in various liver diseases in man.
通过楔形活检或针吸活检获取的八份人类肝脏组织样本,用0.5% Triton X - 100进行提取,然后通过扫描电子显微镜和免疫电子显微镜进行研究。楔形活检标本用去污剂溶液灌注。针吸活检标本迅速冷冻并裂开,然后将裂开的组织浸入去污剂溶液中。肝细胞的三维丝状网络得以显现。在肝细胞胞质内观察到由中间丝和微丝组成的致密网络。与楔形活检相比,这些细丝在提取前经过快速冷冻和裂开处理的针吸活检中保存得更好。在经过快速冷冻处理的肝细胞中,细胞骨架细丝数量的变化不太明显。结论是,冷冻裂开是研究人类各种肝脏疾病细胞骨架病理学的最有利方法。