Comprehensive SummaryOrganic difluoroboron complexes are a kind of potential platforms for a wide range of applications owing to their excellent photophysical properties. Herein, we have explored a simple and direct synthesis methodology for a library of N,O‐bidentate difluoroboron complexes from quinoxalin‐2(1H)‐ones and ketones in one shot. The photophysical properties of the generated complexes were evaluated and the application potential of these compounds on subcellular imaging was also explored.