Leonard E J, Yoshimura T, Skeel A, Goodwin R
Laboratory of Immunobiology, National Cancer Institute, FCRDC, Frederick, MD 21702.
Biotechniques. 1990 Dec;9(6):684, 686, 688-9.
A simple multiwell chamber is described that can be used to prepare randomly distributed cells on a microscope slide, suitable for morphological identification and differential counting. To the eight wells of the chamber are added 50-microliter volumes of cell suspension at concentrations of 10(3)-10(6) cells/ml. As the cells settle, fluid is slowly wicked away by a damp filter paper sandwiched between the microscope slide and the acrylic top plate of the multiwell chamber. Within 20-40 minutes, the cell monolayers on the slide are completely dry. The combined settling and bulk fluid removal results in a distribution of adherent cells that are sufficiently spread to exhibit excellent morphology after staining. If the chamber is centrifuged for 30 seconds at 50x g immediately after addition of cells, recovery of cells in the monolayer is virtually 100%, and as few as 50 input cells per 50 microliters can be detected. Agreement between predicted and observed differential counts of cell mixtures indicates that cells in the monolayer were distributed randomly.
描述了一种简单的多孔腔室，可用于在显微镜载玻片上制备随机分布的细胞，适用于形态学鉴定和分类计数。向该腔室的八个孔中加入浓度为10³ - 10⁶个细胞/毫升的50微升细胞悬液。当细胞沉降时，夹在显微镜载玻片和多孔腔室丙烯酸顶板之间的湿滤纸会缓慢吸走液体。在20 - 40分钟内，载玻片上的细胞单层完全干燥。沉降和大量液体去除相结合，使得贴壁细胞分布充分散开，染色后呈现出极佳的形态。如果在加入细胞后立即以50×g离心腔室30秒，单层细胞的回收率几乎为100%，每50微升中低至50个输入细胞都能被检测到。细胞混合物预测和观察到的分类计数之间的一致性表明单层中的细胞是随机分布的。
