Spriggs T L, Wynne-Evans D
J Microsc. 1976 May;107(1):35-46. doi: 10.1111/j.1365-2818.1976.tb02421.x.
Thin sections of unfixed liver, fast-frozen without cryoprotectants, have been cut using conditions under which momentary thawing of the sections is unlikely to occur. A transfer stage which facilitates this procedure is described. Sections show hole damage probably due to ice-crystal formation during the freezing process and have well defined edges, but despite hole damage, some morphological features of the cell are discernible. Presumptive mitochondria appear smaller in frozen sections than in conventional Araldite sections. Sections devoid of hole damage have indistinct edges and are presumed to have undergone transient thawing. Carbon coating of freeze-dried sections to exclude atmospheric moisture during transference of sections to the electron microscope (EM) appear unnecessary as regards preservation of morphological structure. The results are discussed in relation to the limitations of the method and the potential value of the technique.
未固定的肝脏薄切片,在未使用冷冻保护剂的情况下快速冷冻,切片时采用不太可能出现瞬间解冻的条件。描述了一种便于此操作的转移台。切片显示出可能是由于冷冻过程中冰晶形成导致的孔洞损伤,边缘清晰,但尽管有孔洞损伤,细胞的一些形态特征仍可辨别。推测冷冻切片中的线粒体比传统环氧树脂包埋切片中的线粒体小。没有孔洞损伤的切片边缘不清晰,推测经历了短暂解冻。就形态结构的保存而言,冷冻干燥切片在转移至电子显微镜(EM)过程中进行碳涂层以排除大气水分似乎没有必要。结合该方法的局限性和该技术的潜在价值对结果进行了讨论。