Sawitzky H, Willingale-Theune J, Menzel D
Max-Planck-Institut für Zellbiologie, Rosenhof, Ladenburg, Germany.
Histochem J. 1996 May;28(5):353-60. doi: 10.1007/BF02331398.
By employing a new procedure we have been able to visualize a highly intense actin cytoskeleton in the unicellular green alga Acetabularia acetabulum Silva. The protocol described in this study involves microwave-accelerated simultaneous permeabilization with 10% dimethyl sulphoxide, fixation with 1% glutaraldehyde and incubation with 0.5 microM fluorescein-isothiocyanate-conjugated Phalloidin. Comparison of the images of the actin cytoskeleton of the stalk, as visualized by methods used previously, with those obtained in our own experiments shows that the actin filaments were preserved completely in an excellent condition. The required time for each procedure could be reduced from 12 h for the most commonly used immunofluorescence technique to 35 min. Moreover, it has been possible to observe the actin filament system of hair whorls, rhizoid and tip. Previously, the actin cytoskeleton of these parts of the cell could not be visualized by conventional techniques. It is shown that each region of the cell-stalk, tip, rhizoid and side branches-displays characteristic degrees of actin bundling and regularity of actin alignment.
通过采用一种新方法，我们得以在单细胞绿藻伞藻（Acetabularia acetabulum Silva）中观察到高度密集的肌动蛋白细胞骨架。本研究中描述的实验方案包括用10%二甲基亚砜进行微波加速同步透化处理、用1%戊二醛固定以及用0.5微摩尔异硫氰酸荧光素偶联的鬼笔环肽孵育。将先前使用的方法所观察到的柄部肌动蛋白细胞骨架图像与我们自己实验中获得的图像进行比较，结果表明肌动蛋白丝完全保持在极佳状态。每个步骤所需的时间可以从最常用的免疫荧光技术的12小时减少到35分钟。此外，还能够观察到毛状体、假根和顶端的肌动蛋白丝系统。此前，通过传统技术无法观察到细胞这些部位的肌动蛋白细胞骨架。结果表明，细胞的每个区域——柄部、顶端、假根和侧枝——都呈现出肌动蛋白束集的特征程度和肌动蛋白排列规律。
