Harnessing J-aggregation for dual-color cellular imaging with chromenoquinoline-benzimidazole dyes†

Fluorescence imaging has revolutionized the visualization of cellular structures and biomolecules due to its non-invasive nature and high sensitivity. Chromenoquinoline (CQ)-based dyes offer promising optical properties, yet their widespread application is hindered by aggregation-caused quenching (ACQ). In contrast, J-aggregates, characterized by distinctive photophysical properties, present a solution to ACQ. Here, we introduce a novel platform employing chromenoquinoline-benzimidazole (CQ-BI) dyes, capable of forming J-aggregates, for dual-color cellular imaging. The incorporation of a methyl group into the benzimidazole moiety enhances J-aggregate formation, leading to robust emission in both dilute solutions and aggregated states. Our study demonstrates that methyl moiety-modified CQ-BI derivatives enable simultaneous imaging of mitochondria and lipid droplets in living cells. This work underscores the potential of CQ-BI dyes for dual-channel fluorescence imaging, leveraging the unique properties of J-aggregation.