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Peer-reviewedCC BYJournal of biomedical optics

Near-infrared fluorescence lifetime imaging: fluorophores, methods, and applications.

  • Nanxue Yuan1
  • Catherine Sherry2
  • Navid Ibtehaj Nizam3
  • Saif Ragab1
  • Luis Chavez1
  • Vikas Pandey1
  • Amit Verma2
  • Colleen N Scott4
  • Margarida M Barroso2
  • Xavier Intes1
4 affiliations
  1. 1Rensselaer Polytechnic Institute, Center for Modeling, Simulation and Imaging in Medicine, Troy, New York, United States.
  2. 2Albany Medical College, Department of Molecular and Cellular Physiology, Albany, New York, United States.
  3. 3West Chester University, Department of Biomedical Engineering, West Chester, Pennsylvania, United States.
  4. 4Mississippi State University, Department of Chemistry, Starkville, Mississippi, United States.
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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.Results

Reported NIR fluorophore lifetimes span tens of picoseconds to over 100 ns and depend on molecular structure, solvent, binding state, and aggregation. Multiplexing requires reproducible, distinguishable decay signatures; sensing requires selective, calibrated lifetime responses; and FRET requires reliable donor-only references. Uncorrected instrumental effects, tissue photon migration, and inappropriate analysis assumptions can distort lifetime estimates. Incomplete characterization and inconsistent reporting further limit reproducibility and comparison across studies.Conclusions

Quantitative NIR FLI requires precise probe characterization, standardized measurement validation, and application-specific performance assessment. We provide practical recommendations for probe selection, lifetime contrast interpretation, and clinical utility evaluation, establishing a framework for reproducible NIR FLI from mechanistic cell studies to image-guided interventions.

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Cite

BibTeXRIS
BibTeX
@article{yuan2026nearinfrared,
  title = {{Near-infrared fluorescence lifetime imaging: fluorophores, methods, and applications.}},
  author = {Nanxue Yuan and Catherine Sherry and Navid Ibtehaj Nizam and Saif Ragab and Luis Chavez and Vikas Pandey and Amit Verma and Colleen N Scott and Margarida M Barroso and Xavier Intes},
  journal = {Journal of biomedical optics},
  year = {2026},
  month = oct,
  doi = {10.1117/1.jbo.31.10.100901},
  url = {https://europepmc.org/article/MED/42922939},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Near-infrared fluorescence lifetime imaging: fluorophores, methods, and applications.
AU  - Nanxue Yuan
AU  - Catherine Sherry
AU  - Navid Ibtehaj Nizam
AU  - Saif Ragab
AU  - Luis Chavez
AU  - Vikas Pandey
AU  - Amit Verma
AU  - Colleen N Scott
AU  - Margarida M Barroso
AU  - Xavier Intes
PY  - 2026
DA  - 2026/10/21
JO  - Journal of biomedical optics
DO  - 10.1117/1.jbo.31.10.100901
UR  - https://europepmc.org/article/MED/42922939
AB  - 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.Results  Reported NIR fluorophore lifetimes span tens of picoseconds to over 100 ns and depend on molecular structure, solvent, binding state, and aggregation. Multiplexing requires reproducible, distinguishable decay signatures; sensing requires selective, calibrated lifetime responses; and FRET requires reliable donor-only references. Uncorrected instrumental effects, tissue photon migration, and inappropriate analysis assumptions can distort lifetime estimates. Incomplete characterization and inconsistent reporting further limit reproducibility and comparison across studies.Conclusions  Quantitative NIR FLI requires precise probe characterization, standardized measurement validation, and application-specific performance assessment. We provide practical recommendations for probe selection, lifetime contrast interpretation, and clinical utility evaluation, establishing a framework for reproducible NIR FLI from mechanistic cell studies to image-guided interventions.
SN  - 1083-3668
SN  - 1560-2281
ER  -