Simultaneous recording of multiple cellular events by FRET
Simultaneous recording of multiple cellular events by FRET
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DOI：
10.1021/cb700247q
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发表时间：
2008-03-01
影响因子：
4
通讯作者：
Schultz, Carsten
中科院分区：
文献类型：
--
作者：
Piljic, Alen;Schultz, Carsten
许多传感器用于测量细胞内事件的功能是基于Forster共振能量转移(FRET)的。在这里，我们首次通过空间和光谱分辨率演示了多个比率FRET传感器的并行使用。我们通过钙/钙调蛋白依赖性蛋白激酶IIα的胞浆传感器、蛋白激酶C的膜结合传感器和基于Annexin A4的转位FRET探针监测了三个钙依赖的信号事件。这种多参数成像方法可以深入了解单个细胞内细胞事件的精确时间，从而提供了比单参数协议更大的优势。这种类型的成像可能会对未来的高含量细胞分析和筛选工作起到重要作用。
The function of many sensors for measuring intracellular events is based on Forster resonance energy transfer (FRET). Here we demonstrate for the first time the use of multiple ratiometric FRET sensors in parallel through spatial and spectral resolution. We monitored three calcium-dependent signaling events by a cytosolic sensor for calcium/calmodulin-dependent protein kinase II alpha, a membrane-bound sensor for protein kinase C, and a translocating FRET probe based on annexin A4. This multiparameter imaging approach gives insight into the precise timing of cellular events within one single cell, thereby providing a major advantage over single-parameter protocols. This type of imaging will likely be important for high content cell analysis and screening efforts in the future.