Observation of changes in the intracellular concentration of calcium ion (Ca2+) is important for study of signaling mechanism.Small chemical fluorescent Ca2+ indicators such as fura-2 and fura-PE3 are important tools for studying changes in the intracellular Ca2+ concentration in primary culture cells which are considered difficult to transfect. In this research, a plasma membrane-permeable clickable Ca2+ indicator �N3-fura-2 AM� was synthesized and introduced into HeLa cells, of which cytosolic macromolecules were non-specifically modified with dibenzylcyclooctyne (DBCO) (Figure 1). We confirmed that intracellular Cu-free click reaction occurred in a living cell and Ca2+ oscillation was observed for longer time (at least 6 hours) compared to conventional methods, fura-2 or fura-PE3. Figure 1  . Schematic illustration of intracellular Cu-free click reaction with N3-fura-2 AM.

Intracellular Cu-free click reaction with a small chemical Ca<sup>2+</sup> indicator

248th National Meeting (2014)

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Intracellular Cu-free click reaction with a small chemical Ca2+ indicator

Intracellular Cu-free click reaction with a small chemical Ca²⁺ indicator