Fluorescent-Based Quantitative Measurements of Signal Transduction in Single Cells
Fluorescent-Based Quantitative Measurements of Signal Transduction in Single Cells
复制标题
基于荧光的单细胞信号转导定量测量
DOI：
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
M. Peter
中科院分区：
文献类型：
--
作者：
Serge Pelet;M. Peter
芽殖酵母（Saccharomycescerevisiae）已被广泛用作研究基本生物过程的模型系统。在过去的几十年中，遗传和生物化学方法已经允许揭示参与许多信号通路的关键组分。通常，大多数技术测量细胞群体的平均行为，从而错过重要的细胞间变化。随着荧光蛋白研究的进展，为定量研究单个活细胞中信号传导的动力学开辟了新的途径。在本章中，我们描述了几种基于荧光测量的技术来量化生物途径的激活。流式细胞术允许快速定量大量单细胞的总荧光。相比之下，显微镜的通量较低，但可以以高时间分辨率跟踪亚细胞分辨率的蛋白质定位。最后，先进的功能成像技术，如FRET和FCS提供了直接可视化蛋白质复合物的形成或定量蛋白质在体内的活性的可能性。总之，这些技术提出了强大的新方法来研究细胞信号，并将大大增加我们对芽殖酵母及其他细胞中信号网络调控的理解。
Budding yeast (Saccharomyces cerevisiae) has been widely used as a model system to study fundamental biological processes. Genetic and biochemical approaches have allowed in the last decades to uncover the key components involved in many signaling pathways. Generally, most techniques measure the average behavior of a population of cells, and thus miss important cell-to-cell variations. With the recent progress in fluorescent proteins, new avenues have been opened to quantitatively study the dynamics of signaling in single living cells. In this chapter, we describe several techniques based on fluorescence measurements to quantify the activation of biological pathways. Flow cytometry allows for rapid quantification of the total fluorescence of a large number of single cells. In contrast, microscopy offers a lower throughput but allows to follow with a high temporal resolution the localization of proteins at sub-cellular resolution. Finally, advanced functional imaging techniques such as FRET and FCS offer the possibility to directly visualize the formation of protein complexes or to quantify the activity of proteins in vivo. Together these techniques present powerful new approaches to study cellular signaling and will greatly increase our understanding of the regulation of signaling networks in budding yeast and beyond.
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影响因子：
3.4
作者：
Schwille, P;Haupts, U;Webb, WW
通讯作者：
Webb, WW
影响因子：
2.2
作者：
George, TC;Fanning, SL;Lynch, DH
通讯作者：
Lynch, DH
DOI：
10.1093/bfgp/elm011
发表时间：
2007-06-01
期刊：
Briefings in Functional Genomics & Proteomics
影响因子：
--
作者：
Nolan, John P.;Yang, Loretta
通讯作者：
Yang, Loretta
DOI：
10.1002/cyto.990030402
发表时间：
1983-01-01
期刊：
CYTOMETRY
影响因子：
--
作者：
SHAPIRO, HM
通讯作者：
SHAPIRO, HM
影响因子：
3.3
作者：
Gasch, AP;Spellman, PT;Brown, PO
通讯作者：
Brown, PO