A one-step imaging assay to monitor cell cycle state and apoptosis in mammalian cells.
A one-step imaging assay to monitor cell cycle state and apoptosis in mammalian cells.
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一种监测哺乳动物细胞细胞周期状态和细胞凋亡的一步成像测定。
DOI：
10.1002/9780470559277.ch130140
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发表时间：
2014
影响因子：
--
通讯作者：
Tang,Yangzhong
中科院分区：
文献类型：
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作者：
Tang,Yangzhong
高内容筛选（HCS;具有多个标记的荧光显微术，随后进行自动图像分析）由于其揭示了关于药物反应的丰富信息而在药物发现中越来越受欢迎。它在研究抗有丝分裂药物反应中特别有用，因为有丝分裂停滞提供了活性生物标志物。探测有丝分裂停滞和下游凋亡反应的一种常规方法是在基于细胞的成像测定中使用有丝分裂和凋亡特异性抗体。然而，弱附着的细胞，特别是死细胞，在抗体标记步骤中大部分被洗掉。在这里，我们报告了一种快速方便的一步细胞成像检测方法，可以准确测量哺乳动物细胞的细胞周期状态和细胞凋亡。该检测使用三种荧光染料对活细胞进行染色，无需洗涤，并且在活细胞标记后可固定。与基于抗体的方法相比，该测定更快，更具成本效益，并产生更准确的剂量反应结果。方案Chem.Biol.6：1 - 5.© 2014年由John Wiley & Sons，Inc.
High‐content screening (HCS; fluorescence microscopy with multiple markers followed by automated image analysis) is gaining popularity in drug discovery due to the rich information it reveals about drug responses. It is particularly useful in studying anti‐mitotic drug responses since mitotic arrest provides an activity biomarker. One conventional way to probe mitotic arrest and downstream apoptosis response is to use mitosis‐ and apoptosis‐specific antibodies in cell‐based imaging assays. However, weakly attached cells, especially dead cells, are mostly washed out during antibody labeling steps. Here, we report a rapid and convenient one‐step cell‐imaging assay that accurately measures cell‐cycle state and apoptosis in mammalian cells. The assay uses three fluorescent dyes to stain living cells, involves no wash, and is fixable after live‐cell labeling. Compared to the antibody‐based method, this assay is quicker, more cost‐effective, and yields more accurate dose‐response results.Curr. Protoc. Chem. Biol. 6:1‐5. © 2014 by John Wiley & Sons, Inc.