Modha K, Whiteside J P, Spier R E
Department of Microbiology, University of Leicester, UK.
Cytotechnology. 1993;13(3):227-32. doi: 10.1007/BF00749819.
A colorimetric assay utilising Neutral Red (C.I. 50040), a nuclear stain, was developed to determine the cellular viability of hybridoma cells in microtitre plates. A linear correlation (r = 0.99) was found to exist between the uptake of Neutral Red by viable cells and the viable cell count determined by Trypan blue exclusion test. The linearity stretched over the range of cell concentrations normal in batch cultures (2-30 x 10(4)/0.2 ml) with as little as +/- 6% intra-plate well-to-well variation and +/- 10.2% inter-assay variation. Microscopical examinations of viable hybridoma cells stained with Neutral Red showed that it was located in the nucleus. The possible bifunctional activity of the Neutral Red assay as a test for cellular viability and estimating the DNA content of hybridoma cells is discussed along with its application in a drug screening programme.
开发了一种利用核染色剂中性红（C.I. 50040）的比色测定法，以确定微量滴定板中杂交瘤细胞的细胞活力。发现活细胞摄取中性红的量与通过台盼蓝排斥试验确定的活细胞计数之间存在线性相关性（r = 0.99）。线性范围涵盖分批培养中正常的细胞浓度范围（2 - 30×10⁴/0.2 ml），板内孔间变化小至±6%，测定间变化为±10.2%。用中性红染色的活杂交瘤细胞的显微镜检查表明，中性红位于细胞核中。讨论了中性红测定法作为细胞活力测试和估计杂交瘤细胞DNA含量的可能双功能活性及其在药物筛选程序中的应用。
