Fluorescence-based cell viability screening assays using water-soluble oxygen probes
Fluorescence-based cell viability screening assays using water-soluble oxygen probes
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DOI：
10.1177/1087057103008003004
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发表时间：
2003-06-01
影响因子：
--
通讯作者：
Papkovsky, DB
中科院分区：
文献类型：
--
作者：
Hynes, J;Floyd, S;Papkovsky, DB
描述了一种简单的基于发光的方法，用于筛选哺乳动物细胞的活性，该方法基于通过磷光水溶性氧探针监测细胞呼吸的方法，该探针通过改变发射强度和寿命来响应溶解氧浓度的变化。在低浓度(0.3微米到0.5毫微米)下将探针加入到每个样品中，每个样品包含标准96孔板的孔中的细胞培养。通过在每个样品上涂上一层矿物油，然后在即时或时间分辨的荧光板读取器上监测磷光信号，开始分析氧气消耗。摄氧率可以根据磷光动力学变化(初始斜率)来确定，并与细胞数量(对于FL5.12淋巴母细胞系为10(5)到10(7)个/毫升)、细胞活力或药物/效应作用相关。与现有的基于微孔板的细胞活力分析方法相比，该方法是一种非侵入性的、更简单、更可靠、更具成本效益的检测方法；与现有仪器兼容；并允许对细胞活力进行高通量分析。(《生物分子筛选杂志》2003：264-272)
A simple luminescence-based assay for screening the viability of mammalian cells is described, based on the monitoring of cell respiration by means of a phosphorescent water-soluble oxygen probe that responds to changes in the concentration of dissolved oxygen by changing its emission intensity and lifetime. The probe was added at low concentrations (0.3 muM to 0.5 nM) to each sample containing a culture of cells in the wells of a standard 96-well plate. Analysis of oxygen consumption was initiated by applying a layer of mineral oil on top of each sample followed by monitoring of the phosphorescent signal on a prompt or time-resolved fluorescence plate reader. Rates of oxygen uptake could be determined on the basis of kinetic changes of the phosphorescence (initial slopes) and correlated with cell numbers (10(5) to 10(7) cells/mL for FL5.12 lymphoblastic cell line), cell viability, or drug/effector action using appropriate control samples. The assay is cell noninvasive, more simple, robust, and cost-effective than existing microplate-based cell viability assays; is compatible with existing instrumentation; and allows for high-throughput analysis of cell viability. (Journal of Biomolecular Screening 2003:264-272).