Safa A R, Tseng M T
Stain Technol. 1982 Mar;57(2):107-12. doi: 10.3109/10520298209066537.
The closeness of wells in multiwell tissue culture plates makes it difficult to remove individual ones without distributing the adjacent wells. Moreover, critical point drying frequently introduces artifact into the culture monolayer grown on plastic substrate. These problems make it difficult to process such cultures for scanning electron microscopy. However, for cell kinetic and correlative morphological studies on primary cultures derived from 7,12-dimethylbenz(alpha)anthracene(DMBA)-induced mammary tumors, we have found that Falcon 24-well tissue culture plates are excellent for maintenance of cells and are convenient to use. By plating the cells in alternating, diagonally disposed wells and while the cells are still in the buffer, individual wells can be cut from a multiwell plate without disturbing the cells in adjacent wells. The isolated wells can be used to carry out scanning electron microscope preparation. The height of the well is reduced with a scalpel prior to critical point drying; the remaining well is useful as a handle in mounting the dried sample to stubs for subsequent coating and viewing. Critical point drying and coating of monolayer samples on Falcon plastic are described. The results do not suggest that any artifacts are introduced in our preparations.
多孔组织培养板中各孔间距较近，要移除单个孔而不影响相邻孔很困难。此外，临界点干燥常常会给生长在塑料基质上的培养单层细胞引入假象。这些问题使得对这类培养物进行扫描电子显微镜观察的处理变得困难。然而，对于源自7,12 - 二甲基苯并(α)蒽（DMBA）诱导的乳腺肿瘤的原代培养细胞进行细胞动力学和相关形态学研究时，我们发现Falcon 24孔组织培养板非常适合细胞培养，且使用方便。通过将细胞接种在交替对角排列的孔中，并且在细胞仍处于缓冲液中时，就可以从多孔板中切割出单个孔，而不会干扰相邻孔中的细胞。分离出的孔可用于进行扫描电子显微镜样品制备。在临界点干燥之前，用手术刀降低孔的高度；剩余的孔在将干燥后的样品安装到短柱上以便后续镀膜和观察时可作为手柄使用。本文描述了在Falcon塑料上对单层样品进行临界点干燥和镀膜的方法。结果表明我们的制备过程未引入任何假象。
