Multicolor monitoring of cellular organelles by single wavelength excitation to visualize the mitophagy process

Multiplexed cellular organelle imaging using single wavelength excitation is highly desirable for unravelling cellular functions but remains challenging. This requires the design of organelle specific fluorophores with distinct emission but similar absorption. Herein, we present two unique aggregation-induced emission (AIE) probes to track mitochondria and lysosomes simultaneously with emission colors that can be distinguished from that of the nucleus stain Hoechst 33342 upon single wavelength excitation. Compared to conventional organelle stains, the two AIE probes have larger Stokes shifts and higher photostability, which endow them with the capability to monitor bioprocesses, such as mitophagy with strong and sustained fluorescent signals. Moreover, both probes can also stain intracellular organelles in zebrafish larvae with good cell-penetrating capabilities, showing their great potential to monitor bioprocesses in vivo.