Multichannel fluorescent probes for discriminative detection

Multichannel fluorescent probes have emerged as powerful molecular tools for simultaneously visualizing and quantifying multiple biomolecule within living systems. Compared with conventional single- or dual-channel probes, multichannel designs integrate several specific reactive sites or spectrally tunable units within a single molecular scaffold, thereby enabling precise discrimination of structurally similar or coexisting analytes. In recent years, the applications of multichannel fluorescent probes have expanded beyond small-molecule detection (such as cysteine, homocysteine, glutathione, hydrogen sulfide, and reactive oxygen species) to more complex biological processes, including organelle communication, polarity and viscosity mapping, and multiparametric disease diagnosis. These innovations have markedly improved spatiotemporal resolution, detection accuracy, and biocompatibility, providing new opportunities to explore cellular microenvironments and disease mechanisms. This review systematically summarizes the recent advances in the design principles, sensing mechanisms at the molecular level, and representative applications of multichannel fluorescent probes, emphasizing their unique advantages and promising potential in disease diagnostics, and precision medicine.