Bridging the translation gap: how fluorophore design and imaging innovations will shape next-generation point-of-care diagnostics.

Fluorescence-based point-of-care testing (POCT) holds transformative potential for global health, molecular diagnostics, and personalized medicine, yet the path from laboratory prototype to clinical deployment remains obstructed by unresolved gaps in probe chemistry, imaging hardware, data analytics, and validation. This critical review examines the design and integration innovations most likely to close that gap. We assess near-infrared (NIR) fluorophores, nanomaterial probes including quantum dots (QDs), carbon dots and aggregation-induced emission (AIE) luminogens, and multimodal fluorescence/surface-enhanced Raman scattering (SERS) systems, evaluating trade-offs among brightness, photostability, multiplexing capacity, and biocompatibility. We analyze strategies that improve signal-to-noise ratio (SNR) in complex biological matrices, including activatable and ratiometric probe designs, plasmonic amplification, and on-strip sample conditioning. Integration into paper-based and microfluidic platforms is discussed alongside the practical challenges of smartphone-based readers, particularly illumination control and cross-device standardization. We further address constraints specific to nucleic-acid POCT, notably thermal management, and evaluate hybrid paper-microfluidic architectures as a practical workaround. Drawing these threads together, we propose a translational framework built on standardized optical and analytical pipelines, robust background suppression, reproducible substrate fabrication, and multi-site clinical validation. The goal is to provide actionable design and validation principles that accelerate the adoption of fluorescence POCT where it is most needed.