CalQuo: automated, simultaneous single-cell and population-level quantification of global intracellular Ca2+ responses.
CalQuo: automated, simultaneous single-cell and population-level quantification of global intracellular Ca2+ responses.
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DOI：
10.1038/srep16487
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发表时间：
2015-11-13
影响因子：
4.6
通讯作者：
Eggeling C
中科院分区：
文献类型：
--
作者：
Fritzsche M;Fernandes RA;Colin-York H;Santos AM;Lee SF;Lagerholm BC;Davis SJ;Eggeling C
用荧光钙指示剂染料检测细胞内钙信号通常与显微镜技术相结合，以跟踪非兴奋细胞（包括淋巴细胞）的活化状态。然而，在单细胞水平和同时在大型合奏的全球细胞内钙反应的分析还有待自动化。在这里，我们提出了一个新的软件包，CalQuo（钙定量），它允许自动分析和同时监测全球荧光钙标记物为基础的信号反应，在多达1000个单细胞的实验，在亚秒到秒的时间分辨率。CalQuo定量响应细胞的数量和分数、钙信号传导的时间依赖性，并提供全局和个体钙报告荧光强度分布。我们证明了新方法的实用性，通过比较基于钙的信号转导反应的遗传操作的人淋巴细胞系。
Detecting intracellular calcium signaling with fluorescent calcium indicator dyes is often coupled with microscopy techniques to follow the activation state of non-excitable cells, including lymphocytes. However, the analysis of global intracellular calcium responses both at the single-cell level and in large ensembles simultaneously has yet to be automated. Here, we present a new software package, CalQuo (Calcium Quantification), which allows the automated analysis and simultaneous monitoring of global fluorescent calcium reporter-based signaling responses in up to 1000 single cells per experiment, at temporal resolutions of sub-seconds to seconds. CalQuo quantifies the number and fraction of responding cells, the temporal dependence of calcium signaling and provides global and individual calcium-reporter fluorescence intensity profiles. We demonstrate the utility of the new method by comparing the calcium-based signaling responses of genetically manipulated human lymphocytic cell lines.