Journal of biomedical optics
Near-infrared fluorescence lifetime imaging: fluorophores, methods, and applications
By Yuan N, Sherry C, Nizam NI and 7 others.
Abstract
Significance
Near-infrared (NIR) fluorescence lifetime imaging (FLI) provides molecular and functional contrast that encodes local biochemical and biophysical environments beyond what fluorescence intensity alone can reveal. Realizing this potential across biological imaging and emerging clinical applications requires rigorous fluorophore characterization and a critical assessment of the photophysics, measurement capabilities, and conditions governing reliable lifetime interpretation.
Aim
We comprehensively survey the contemporary NIR FLI landscape, spanning fluorophore photophysics, imaging instrumentation, lifetime analysis, and biomedical applications, to identify application-specific criteria for fluorophore selection, quantitative measurement, and translational validation. Approach: Within the NIR range, we critically compare organic dyes, fluorescent proteins, inorganic fluorophores, and endogenous emitters, integrating photophysical evidence with acquisition and analysis methods for multiplexing, Förster resonance energy transfer (FRET), environmental sensing, and clinical imaging.