Academic Journal
Multiphoton imaging coupled with steady-state and time-resolved fluorescence spectroscopy reveals protein-bound FAD in femtosecond-laser-injured regenerating muscle.
| Τίτλος: | Multiphoton imaging coupled with steady-state and time-resolved fluorescence spectroscopy reveals protein-bound FAD in femtosecond-laser-injured regenerating muscle. |
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| Συγγραφείς: | Wang ZW; Department of Applied Chemistry, National Yang Ming Chiao Tung University, 1001 University Road, Hsinchu 300, Taiwan; Center for Emergent Functional Matter Science, National Yang Ming Chiao Tung University, 1001 University Road, Hsinchu 300, Taiwan., Lin HJ; Department of Applied Chemistry, National Yang Ming Chiao Tung University, 1001 University Road, Hsinchu 300, Taiwan; Center for Emergent Functional Matter Science, National Yang Ming Chiao Tung University, 1001 University Road, Hsinchu 300, Taiwan., Fathi A; Department of Applied Chemistry, National Yang Ming Chiao Tung University, 1001 University Road, Hsinchu 300, Taiwan; Center for Emergent Functional Matter Science, National Yang Ming Chiao Tung University, 1001 University Road, Hsinchu 300, Taiwan., Liau I; Department of Applied Chemistry, National Yang Ming Chiao Tung University, 1001 University Road, Hsinchu 300, Taiwan; Center for Emergent Functional Matter Science, National Yang Ming Chiao Tung University, 1001 University Road, Hsinchu 300, Taiwan. Electronic address: ianliau@nycu.edu.tw. |
| Πηγή: | Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy [Spectrochim Acta A Mol Biomol Spectrosc] 2026 Apr 05; Vol. 350, pp. 127403. Date of Electronic Publication: 2025 Dec 29. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Elsevier Country of Publication: England NLM ID: 9602533 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-3557 (Electronic) Linking ISSN: 13861425 NLM ISO Abbreviation: Spectrochim Acta A Mol Biomol Spectrosc Subsets: MEDLINE |
| Imprint Name(s): | Publication: Original Publication: [Kidlington, Oxford, U.K. ; Tarrytown, NY] : Pergamon, c1994- |
| Ιατρικοί όροι (MeSH): | Flavin-Adenine Dinucleotide*/metabolism , Muscle, Skeletal*/injuries , Muscle, Skeletal*/metabolism , Muscle, Skeletal*/physiology , Microscopy, Fluorescence, Multiphoton*/methods , Regeneration* , Lasers*, Spectrometry, Fluorescence/methods ; Reactive Oxygen Species/metabolism ; Sarcomeres/metabolism ; Animals ; Zebrafish ; Oxidation-Reduction |
| Περίληψη: | Regeneration of skeletal muscle involves tightly coupled structural and redox processes that remain difficult to characterize in vivo or in situ with molecular specificity. Here, we introduce a label-free spectroscopic imaging framework that integrates multiphoton imaging-including second-harmonic generation (SHG) and two-photon-excited fluorescence (TPEF)-with steady-state and time-resolved fluorescence spectroscopy to visualize sarcomere remodeling and redox dynamics in living zebrafish muscle. SHG imaging revealed a transient disruption and gradual restoration of sarcomeric order, indicating an asynchronous, sequential reassembly of myofibrillar structures following localized femtosecond laser ablation. In parallel, TPEF imaging detected a pronounced and spatially confined increase in intrinsic emission at the injury site. Single-photon fluorescence spectroscopy and lifetime analysis identified this emission as protein-bound, oxidized flavin adenine dinucleotide (FAD), matching the photophysical signature of mitochondrial flavoproteins and revealing localized redox activation during the early phase of repair. Complementary reactive‑oxygen-species (ROS) imaging confirmed localized oxidative stress associated with elevated electron-transport activity, linking mitochondrial metabolism to structural recovery. Together, these multimodal observations establish a quantitative, label-free framework for correlating mitochondrial FAD-mediated redox activity with sarcomeric remodeling, establishing a versatile optical platform for investigating flavin photophysics, oxidative metabolism, and regenerative physiology in living muscle tissues. (Copyright © 2025 Elsevier B.V. All rights reserved.) |
| Competing Interests: | Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. |
| Contributed Indexing: | Keywords: Femtosecond-laser ablation; Flavin adenine dinucleotide (FAD); Fluorescence lifetime spectroscopy; Mitochondrial redox metabolism; Second-harmonic generation (SHG); Skeletal muscle regeneration; Two-photon excited fluorescence (TPEF) |
| Substance Nomenclature: | 146-14-5 (Flavin-Adenine Dinucleotide) 0 (Reactive Oxygen Species) |
| Entry Date(s): | Date Created: 20260101 Date Completed: 20260129 Latest Revision: 20260129 |
| Update Code: | 20260130 |
| DOI: | 10.1016/j.saa.2025.127403 |
| PMID: | 41478074 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1873-3557 |
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| DOI: | 10.1016/j.saa.2025.127403 |