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A novel styrylbenzimidazolium dye containing a maleimide group 5 was synthesized and characterized using proton nuclear magnetic resonance spectroscopy and mass spectrometry. The photophysical properties [ultraviolet-visible (UV-vis) light absorption and fluorescence spectra, fluorescence quantum yield, and fluorescence lifetime] were investigated. Spectroscopic characterization of the novel styrylbenzimidazolium-based dye under various conditions is presented and its usefulness to detect biothiols proved. The addition of biothiols [l-cy