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细胞与聚赖氨酸包被表面的黏附。在电子显微镜中的应用。

Adhesion of cells to surfaces coated with polylysine. Applications to electron microscopy.

作者信息

Mazia D, Schatten G, Sale W

出版信息

J Cell Biol. 1975 Jul;66(1):198-200. doi: 10.1083/jcb.66.1.198.

Abstract

Cells of many kinds adhere firmly to glass or plastic surfaces which have been pretreated with polylysine. The attachment takes place as soon as the cells make contact with the surfaces, and the flattening of the cells against the surfaces is quite rapid. Cells which do not normally adhere to solid surfaces, such as sea urchin eggs, attach as well as cells which normally do so, such as amebas or mammalian cells in culture. The adhesion is interpreted simply as the interaction between the polyanionic cell surfaces and the polycationic layer of adsorbed polylysine. The attachment of cells to the polylysine-treated surfaces can be exploited for a variety of experimental manipulations. In the preparation of samples for scanning or transmission electron microscopy, the living material may first be attached to a polylysine-coated plate or grid, subjected to some experimental treatment (fertilization of an egg, for example), then transferred rapidly to fixative and further passed through processing for observation; each step involves only the transfer of the plate or grid from one container to the next. The cells are not detached. The adhesion of the cell may be so firm that the body of the cell may be sheared away, leaving attached a patch of cell surface, face up, for observation of its inner aspect. For example, one may observe secretory vesicles on the inner face of the surface (3) or may study the association of filaments with the inner surface (Fig. 1). Subcellular structures may attach to the polylysine-coated surfaces. So far, we have found this to be the case for nuclei isolated from sea urchin embryos and for the microtubules of flagella, which are well displayed after the membrane has been disrupted by Triton X-100 (Fig. 2).

摘要

许多种类的细胞会牢固地附着在经过聚赖氨酸预处理的玻璃或塑料表面。一旦细胞与这些表面接触,附着过程就会立即发生,并且细胞会迅速在表面上扁平化。通常不附着于固体表面的细胞,如海参卵,也能像正常附着的细胞一样附着,如培养中的变形虫或哺乳动物细胞。这种黏附被简单地解释为多阴离子细胞表面与吸附的聚赖氨酸的聚阳离子层之间的相互作用。细胞与聚赖氨酸处理过的表面的附着可用于各种实验操作。在制备用于扫描或透射电子显微镜的样品时,生物材料可以首先附着在涂有聚赖氨酸的平板或网格上,进行一些实验处理(例如卵子受精),然后迅速转移到固定剂中,并进一步经过处理以进行观察;每个步骤仅涉及将平板或网格从一个容器转移到另一个容器。细胞不会脱落。细胞的黏附可能非常牢固,以至于细胞体可能被剪切掉,留下一块朝上的细胞表面贴片,用于观察其内部结构。例如,可以观察表面(3)内表面上的分泌小泡,或者研究细丝与内表面的关联(图1)。亚细胞结构可能附着在涂有聚赖氨酸的表面上。到目前为止,我们发现从海胆胚胎中分离出的细胞核以及鞭毛的微管就是这种情况,在用Triton X - 100破坏膜后,它们能很好地显示出来(图2)。

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