We have synthesized and characterized an acceptor-pi-donor-pi-acceptor type pyridinium salt. The salt exhibited yellow emission with large Stokes shift in solution and was successfully excited by NIR irradiation via two-photon absorption process. In addition, it was found that the usage of the salt as a bioimaging agent for HeLa cells visualizes cell death. The combined use of the salt and a commercially available green fluorescent dye for the detection of live cell viability realized the one-shot fluorescence imaging of live and dead HeLa cells under the single blue excitation.