Multiphoton imaging coupled with steady-state and time-resolved fluorescence spectroscopy reveals protein-bound FAD in femtosecond-laser-injured regenerating muscle.

Λεπτομέρειες βιβλιογραφικής εγγραφής
Τίτλος: Multiphoton imaging coupled with steady-state and time-resolved fluorescence spectroscopy reveals protein-bound FAD in femtosecond-laser-injured regenerating muscle.
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: : Amsterdam : Elsevier
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
Περιγραφή
ISSN:1873-3557
DOI:10.1016/j.saa.2025.127403