ArticleProceedings of the National Academy of Sciences of the United States of America1992
Fluorescence lifetime imaging of free and protein-bound NADH.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1992. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 355 papers.
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355 citing papers in PubMed, 783 citations in OpenAlex.
- Emerging trends of fluorescence lifetime imaging microscopy (FLIM): advances, challenges, and prospects.Biophysics reports · 2026Article
- Label-free optical biomarker for prostate cancer diagnosis.Molecular medicine (Cambridge, Mass.) · 2026Article
- Autofluorescence lifetime imaging resolves cell heterogeneity within peripheral blood mononuclear cells.Biophotonics discovery · 2026Article
- FLIM quantification of temperature-dependent enzyme-NADH binding dynamics.Biomedical optics express · 2026Article
- Segmentation-guided photon pooling enables robust single-cell analysis and fast fluorescence lifetime imaging microscopy.Journal of biomedical optics · 2026Article
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- Label-free metabolic imaging monitors the fitness of chimeric antigen receptor T cells.Nature biomedical engineering · 2026Article
- Autofluorescence lifetime imaging resolves cell heterogeneity within peripheral blood mononuclear cells.bioRxiv : the preprint server for biology · 2026Article
- An automated image analysis pipeline for wide-field optical redox imaging of patient-derived cancer organoids.Scientific reports · 2026Article
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- Real-time, high-resolution metabolic characterization of live bacteria using label-free optical metabolic imaging.NPJ biofilms and microbiomes · 2026Article
- Label-free fluorescence lifetime imaging can distinguish cancer from healthy tissue in spontaneously occurring canine oral tumors.Scientific reports · 2026Article
- Divergent role of CD8 T cells with distinct metabolic phenotypes during curative radio-immunotherapy in hot versus cold tumors.bioRxiv : the preprint server for biology · 2026Article
- A fluorescence lifetime separation approach for FLIM live-cell imaging.Journal of microscopy · 2026Article
- Expanding the multiplexing capability of HIDE probes via fluorescence lifetime imaging microscopy.Methods in enzymology · 2026Article
- Label-free human skin imaging with enhanced molecular contrast via time-resolved fluorescence and advanced phasor analysis.Communications biology · 2025Article
- C1q drives neural stem cell quiescence by regulating cell cycle and metabolism through BAI1.Nature communications · 2025Article
- Fluorescence Lifetime Imaging Techniques-A Review on Principles, Applications and Clinical Relevance.Journal of biophotonics · 2025Review
- Label-free nonlinear microscopy probes cellular metabolism and myelin dynamics in live tissue.Communications biology · 2025Article
295 more citing papers are in PubMed but not listed here.
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We introduce a methodology, fluorescence lifetime imaging (FLIM), in which the contrast depends on the fluorescence lifetime at each point in a two-dimensional image and not on the local concentration and/or intensity of the fluorophore. We used FLIM to create lifetime images of NADH when free in solution and when bound to malate dehydrogenase. This represents a challenging case for lifetime imaging because the NADH decay times are just 0.4 and 1.0 ns in the free and bound states, respectively. In the present apparatus, lifetime images are created from a series of phase-sensitive images obtained with a gain-modulated image intensifier and recorded with a charge-coupled device (CCD) camera. The intensifier gain is modulated at the light-modulation frequency or a harmonic thereof. A series of stationary phase-sensitive images each obtained with various phase shifts of the gain-modulation signal, is used to determine the phase angle or modulation of the emission at each pixel, which is in essence the lifetime image. We also describe am imaging procedure that allows specific decay times to be suppressed, allowing in this case suppression of the emission from either free or bound NADH. Since the fluorescence lifetimes of probes are known to be sensitive to numerous chemical and physical factors such as pH, oxygen, temperature, cations, polarity, and binding to macromolecules, this method allows imaging of the chemical or property of interest in macroscopic and microscopic samples. The concept of FLIM appears to have numerous potential applications in the biosciences.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.