Single molecular multianalyte (Ca2+, Mg2+) fluorescent probe and applications to bioimaging
Single molecular multianalyte (Ca2+, Mg2+) fluorescent probe and applications to bioimaging
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DOI：
10.1021/ja0528228
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发表时间：
2005-08-10
影响因子：
15
通讯作者：
Suzuki, K
中科院分区：
文献类型：
--
作者：
Komatsu, H;Miki, T;Suzuki, K
细胞内信号转导依赖于空间和时间的信号传递动力学。为了阐明这些递质分子的相互关系，多色成像已被广泛使用。然而，在一个单元中应用多个指示剂的情况下，指示剂的光谱重叠妨碍了精确的定量分析。此外，这些指标的侵入性（毒性）效应、定位、代谢以及光漂白使情况复杂化。在这里，我们表明，单分子多荧光探针可以克服这些问题。虽然细胞内钙离子作为信号传导剂起着关键作用，而镁离子在许多情况下作为辅因子，但这两种阳离子之间的相关性现在是主要问题。我们设计并合成了Ca 2 +− Mg 2+响应的多荧光探针KCM-1。KCM-1在与Ca ~（2+）络合时显示出光谱蓝移，在与Mg ~（2+）络合时显示出光谱红移。在不同的激发波长下分析数据，同时定量Ca 2+和Mg 2+的浓度。此外，通过使用AM-酯方法，细胞内Ca 2+和Mg 2+浓度同时成像。这种类型的细胞内多分析物成像的单分子多荧光探针首次成功地证明。
Intracellular signal transduction relies on spatial and temporal signal transmitter dynamics. To clarify the correlations of these transmitter molecules, multicolor-imaging has been widely used. However, in the case of applying multiple indicators in a cell, spectral overlap of the indicators prevents accurate quantitative analysis. Moreover, the invasive (toxic) effect, the localization, the metabolism, as well as photobleaching of these indicators complicate the situation. Here, we show that single-molecular multifluorescent probes can overcome these problems. While intracellular calcium plays a critical role as a signal transmitter and magnesium acts as a cofactor in many situations, the correlations between the two cations are now the main issue. We designed and synthesized a Ca2+−Mg2+responsive multifluorescent probe, KCM-1. KCM-1 shows a spectral blue shift upon complexation to Ca2+and a red shift to the presence of Mg2+. With data analyzed at different excitation wavelengths, the concentrations of Ca2+and Mg2+are simultaneously quantified. Furthermore, by using the AM-ester method, intracellular Ca2+and Mg2+concentrations are simultaneously imaged. Such a type of intracellular multiple analyte imaging by a single-molecular multifluorescent probe is successfully demonstrated for the first time.