ArticleScientific reports2017
Multicolor two-photon imaging of endogenous fluorophores in living tissues by wavelength mixing.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 64 papers.
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Who cites it
64 citing papers in PubMed, 135 citations in OpenAlex.
- 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
- Fully 3D-printed endomicroscopic objective for two-photon, multi-wavelength excitation microscopy.Biomedical optics express · 2026Article
- Assessing Intracellular Metabolism of Immune Cells In Situ in Live Zebrafish Larvae by Autofluorescence Lifetime Imaging Microscopy of NAD(P)H and FAD.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Label-free nonlinear microscopy probes cellular metabolism and myelin dynamics in live tissue.Communications biology · 2025Article
- A Non-Invasive Approach to Intracellular Measurement in Solar Lentigo: Investigating Mitochondrial Dysfunction and Senescence Mechanisms Associated with Excessive Melanin Deposition.International journal of molecular sciences · 2025Article
- Visualization of oxygen profile in reconstructed human epidermis by phosphorescence-lifetime imaging microscopy using Ir(III) complex.Scientific reports · 2025Article
- MALDI mass spectrometry imaging of extracellular matrix proteins.The Analyst · 2025Review
- Nonlinear Imaging Detection of Organ Fibrosis in Minute Samples for Early Stage Utilizing Dual-Channel Two-Photon and Second-Harmonic Excitation.Biosensors · 2025Article
- Strong Enhancement of Two-Photon Absorption and Emergence of Unusual Extinction Saturation in Silver Sulfide Quantum Dots Integrated with Gold and Silica Nanostructures.ACS applied materials & interfaces · 2025Article
- Consensus guidelines for cellular label-free optical metabolic imaging: ensuring accuracy and reproducibility in metabolic profiling.Journal of biomedical optics · 2025Article
- Two-photon NAD(P)H-FLIM reveals unperturbed energy metabolism of Ascaris suum larvae, in contrast to host macrophages upon artemisinin derivatives exposure.Scientific reports · 2025Article
- Single cell autofluorescence imaging reveals immediate metabolic shifts of neutrophils with activation across biological systems.Frontiers in immunology · 2025Article
- Endogenous mitochondrial NAD(P)H fluorescence can predict lifespan.Communications biology · 2024Article
- Non-degenerate two-photon imaging of deep rodent cortex using indocyanine green in the water absorption window.Biomedical optics express · 2024Article
- Emerging Functional Connections Between Metabolism and Epigenetic Remodeling in Neural Differentiation.Molecular neurobiology · 2024Review
- Imaging immunometabolismImmunometabolism (Cobham, Surrey) · 2024Article
- Single-sample image-fusion upsampling of fluorescence lifetime images.Science advances · 2024Article
- Non-Degenerate Two-Photon Imaging of Deep Rodent Cortex using Indocyanine Green in the water absorption window.bioRxiv : the preprint server for biology · 2024Article
- Label-free single-cell live imaging reveals fast metabolic switch in T lymphocytes.Molecular biology of the cell · 2024Article
- Probing organoid metabolism using fluorescence lifetime imaging microscopy (FLIM): The next frontier of drug discovery and disease understanding.Advanced drug delivery reviews · 2023Review
4 more citing papers are in PubMed but not listed here.
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Authors and funding
12 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Two-photon imaging of endogenous fluorescence can provide physiological and metabolic information from intact tissues. However, simultaneous imaging of multiple intrinsic fluorophores, such as nicotinamide adenine dinucleotide(phosphate) (NAD(P)H), flavin adenine dinucleotide (FAD) and retinoids in living systems is generally hampered by sequential multi-wavelength excitation resulting in motion artifacts. Here, we report on efficient and simultaneous multicolor two-photon excitation of endogenous fluorophores with absorption spectra spanning the 750-1040 nm range, using wavelength mixing. By using two synchronized pulse trains at 760 and 1041 nm, an additional equivalent two-photon excitation wavelength at 879 nm is generated, and achieves simultaneous excitation of blue, green and red intrinsic fluorophores. This method permits an efficient simultaneous imaging of the metabolic coenzymes NADH and FAD to be implemented with perfect image co-registration, overcoming the difficulties associated with differences in absorption spectra and disparity in concentration. We demonstrate ratiometric redox imaging free of motion artifacts and simultaneous two-photon fluorescence lifetime imaging (FLIM) of NADH and FAD in living tissues. The lifetime gradients of NADH and FAD associated with different cellular metabolic and differentiation states in reconstructed human skin and in the germline of live C. Elegans are thus simultaneously measured. Finally, we present multicolor imaging of endogenous fluorophores and second harmonic generation (SHG) signals during the early stages of Zebrafish embryo development, evidencing fluorescence spectral changes associated with development.
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