Simonson M S, Dunn M J
Department of Medicine, Case Western Reserve University, University Hospitals, Cleveland, Ohio 44106.
Anal Biochem. 1987 Nov 15;167(1):106-12. doi: 10.1016/0003-2697(87)90138-2.
Quantitative studies in cell culture require accurate measurements of cell density and kinetics. We have developed a direct, rapid, and noninvasive method for measuring cell number in monolayer culture. Using computer-aided video microscopy, cell number was measured without detaching or chemically destroying the cells, thereby allowing sequential measurements in the same cell population. Cell number measured by computer-aided microscopy closely correlated with hemocytometer counts and determinations of total cell protein. For high-density monolayers of mesenchymal cells, however, staining was required for accurate counts. Unlike other techniques for measuring cell density, computer-aided microscopy was especially accurate in medium- to low-density cultures (less than 6000 cells/cm2). In addition, we applied this technique to the construction of separate proliferation curves for glomerular mesangial and vascular endothelial cells in coculture. These measurements by cell type in coculture are impossible using conventional methods for determining cell number.
细胞培养中的定量研究需要准确测量细胞密度和动力学。我们开发了一种直接、快速且非侵入性的方法来测量单层培养中的细胞数量。使用计算机辅助视频显微镜，无需分离或化学破坏细胞即可测量细胞数量，从而能够在同一细胞群体中进行连续测量。通过计算机辅助显微镜测量的细胞数量与血细胞计数器计数和总细胞蛋白测定密切相关。然而，对于间充质细胞的高密度单层，需要染色才能进行准确计数。与其他测量细胞密度的技术不同，计算机辅助显微镜在中低密度培养（小于6000个细胞/平方厘米）中特别准确。此外，我们将该技术应用于共培养中肾小球系膜细胞和血管内皮细胞单独增殖曲线的构建。使用传统的细胞数量测定方法无法在共培养中按细胞类型进行这些测量。
