Magnesium-Calcium Multi Fluorescent Probe and Intracellular Appication

细胞内信号转导依赖于信号传递体的时空动态。为了阐明这些相关性，多色成像技术被广泛应用。然而，当在一个细胞光谱中应用多个指标时，这些指标的侵袭效应、定位、代谢和光漂白等会使情况变得复杂。我们认为单分子多荧光探针可以克服这些问题。虽然在许多情况下，细胞内钙作为信号传递者和镁作为辅助因子起着关键作用，但这些相关性是显著的。设计并合成了一种Ca2+-Mg2+荧光探针KCM-1。KCM-1与Ca2+络合呈蓝移，与Mg2+络合呈红移。利用am -酯法将KCM-1成功加载到PC12细胞中，并通过不同激发波长下的数据分析，同时对细胞内Ca2+和Mg2+进行成像。首次证实了单分子多荧光探针对细胞内多种分析物的成像。

Intracellular signal transduction relies on spatial and temporal signal transmitters dynamics. To clarify the correlations, multicolor-imaging has been widely used. However, in the case of applying multiple indicators in a cell spectral overlap of the indicators, invasive effect, localization, metabolism and photobleaching of these indicators complicates the situations. We assumed that single molecular multi fluorescent probes can overcome these problems. While intracellular calcium plays a critical role as signal transmitter and magnesium acts as a cofactor in many situations, these correlations are significant. We designed and synthesized a Ca2+-Mg2+ fluorescent probe, KCM-1. KCM-1 showed a spectral blue shift with complexation to Ca2+ and a red shift to Mg2+. By using AM-ester method, KCM-1 was successfully loaded in PC12 cells and by data analyse at different excitation wavelengths, intracellular Ca2+ and Mg2+ were simultaneously imaged. The intracellular multiple analyte imaging by a single molecular multi fluorescent probe was firstly demonstrated.