Karp R D, Silcox J C, Somlyo A V
J Microsc. 1982 Feb;125(Pt 2):157-65. doi: 10.1111/j.1365-2818.1982.tb00333.x.
A technique is presented showing the use of toluene as a low temperature cement to attach portions of frozen tissues in the desired orientation on to the cryoultramicrotome chuck. Examples of transverse sections of skeletal and vascular smooth muscle are illustrated. Electron probe analysis of these sections indicates that the distribution of elements is maintained. Evidence is presented against through section melting based on sectioning toluene at a temperature just below its melting point (178K). Toluene was sectioned (about 100 nm thickness) successfully at three block temperatures (177, 163 and 113 K) when the knife temperature was by 1 degree or mor colder than the m.p. of toluene. In all cases the toluene sections melted when the knife temperature reached 178K. We conclude that sufficient heat is not generated and/or transferred to the toluene sections during cryosectioning under our conditions to raise the temperature of a 100 nm section by 1 K or more.
本文介绍了一种技术，该技术展示了使用甲苯作为低温黏合剂，将冷冻组织的各部分按所需方向固定在冷冻超薄切片机的载物台上。文中给出了骨骼肌和血管平滑肌横切面的示例。对这些切片的电子探针分析表明元素分布得以保持。基于在略低于甲苯熔点（178K）的温度下对甲苯进行切片，本文提供了反对切片全程融化的证据。当刀温比甲苯熔点低1度或更多时，甲苯在三个样本块温度（177K、163K和113K）下成功切成薄片（约100纳米厚）。在所有情况下，当刀温达到178K时，甲苯切片都会融化。我们得出结论，在我们的条件下进行冷冻切片时，不会产生足够的热量和/或热量不会传递到甲苯切片上，从而使100纳米厚的切片温度升高1K或更多。
