Naib-Majani W, Wohlfarth-Bottermann K E
Eur J Cell Biol. 1985 Jan;36(1):1-7.
The spatial distribution of cytoplasmic actin in endoplasmic drops as well as in plasmodial strands can be demonstrated in cryosections by fluorescently labelled phallotoxins and actin antibodies. Our results on cryosections show an identical fibrillar actin distribution as revealed in semithin sections after conventional fixation and embedding. Thus, it is now possible to apply immunocytochemical analysis to any and all plasmodial stages with or without prior fixation and without using extraction procedures. Consequentially the loss of soluble compounds during processing is avoided. The most protective pretreatment of the living specimens before freezing is a 15 min incubation in 1.5 M sucrose containing 50 mM KCl, 10 mM EGTA and 10 mM PIPES buffer, pH 7.0, at 4 degrees C.
通过荧光标记的鬼笔环肽和肌动蛋白抗体，可在冷冻切片中显示内质滴以及原生质丝中细胞质肌动蛋白的空间分布。我们对冷冻切片的研究结果显示，其纤维状肌动蛋白分布与传统固定和包埋后的半薄切片中显示的相同。因此，现在可以在不进行预先固定和不使用提取程序的情况下，对任何和所有原生质体阶段进行免疫细胞化学分析。从而避免了处理过程中可溶性化合物的损失。在冷冻前对活体标本最具保护作用的预处理方法是，将其在含有50 mM氯化钾、10 mM乙二醇双乙胺四乙酸（EGTA）和10 mM哌嗪-N,N'-双（2-乙磺酸）（PIPES）缓冲液（pH 7.0）的1.5 M蔗糖溶液中于4℃孵育15分钟。
