Magnesium-Calcium Multi Fluorescent Probe and Intracellular Appication

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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.