A high-throughput AO/PI-based cell concentration and viability detection method using the Celigo image cytometry
A high-throughput AO/PI-based cell concentration and viability detection method using the Celigo image cytometry
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DOI：
10.1007/s10616-016-0015-x
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发表时间：
2016-10-01
期刊：
影响因子：
2.2
通讯作者：
Qiu, Jean
中科院分区：
文献类型：
--
作者：
Chan, Leo Li-Ying;Smith, Tim;Qiu, Jean
为了确保正确进行基于细胞的测定，必须确定细胞浓度和活力，以便可以对数据进行标准化以生成有意义且可比较的结果。在免疫肿瘤学、毒理学或生物加工研究中进行的基于细胞的测定通常需要测量多个样品和条件，因此当前使用单个一次性计数载玻片的自动细胞计数器对于高通量筛选测定来说并不实用。近年来，基于板的图像细胞计数系统已被开发用于高通量生物分子筛选分析。在这项工作中，我们使用 Celigo 图像细胞仪演示了一种基于 AO/PI 的高通量细胞浓度和活力方法。首先，我们通过直接与 Cellometer 自动细胞计数器进行比较来验证该方法。接下来，确定细胞浓度动态范围、活力动态范围和一致性。这里描述的高通量 AO/PI 方法可以在 7 分钟内分析 96 孔至 384 孔板样品，这大大减少了基于单个样品的自动细胞计数器所需的时间。此外，该方法可以提高高通量筛选测定的效率，在进行流式细胞术、ELISA 或简单铺板细胞进行细胞培养等测定之前，需要进行多次细胞计数和活力测量。
To ensure cell-based assays are performed properly, both cell concentration and viability have to be determined so that the data can be normalized to generate meaningful and comparable results. Cell-based assays performed in immuno-oncology, toxicology, or bioprocessing research often require measuring of multiple samples and conditions, thus the current automated cell counter that uses single disposable counting slides is not practical for high-throughput screening assays. In the recent years, a plate-based image cytometry system has been developed for high-throughput biomolecular screening assays. In this work, we demonstrate a high-throughput AO/PI-based cell concentration and viability method using the Celigo image cytometer. First, we validate the method by comparing directly to Cellometer automated cell counter. Next, cell concentration dynamic range, viability dynamic range, and consistency are determined. The high-throughput AO/PI method described here allows for 96-well to 384-well plate samples to be analyzed in less than 7 min, which greatly reduces the time required for the single sample-based automated cell counter. In addition, this method can improve the efficiency for high-throughput screening assays, where multiple cell counts and viability measurements are needed prior to performing assays such as flow cytometry, ELISA, or simply plating cells for cell culture.