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Characterisation of NADH fluorescence properties under one-photon excitation with respect to temperature, pH and binding to lactate dehydrogenase

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Title: Characterisation of NADH fluorescence properties under one-photon excitation with respect to temperature, pH and binding to lactate dehydrogenase
Authors: Cannon, T
Lagarto, J
Dyer, B
Garcia Castano, E
Kelly, D
Peters, N
Lyon, A
French, P
Dunsby, C
Item Type: Journal Article
Abstract: Reduced nicotinamide adenine dinucleotide (NADH) is the principal electron donor in glycolysis and oxidative metabolism and is thus recognized as a key biomarker for probing metabolic state. While the fluorescence characteristics of NADH have been investigated extensively, there are discrepancies in the published data due to diverse experimental conditions, instrumentation and microenvironmental parameters that can affect NADH fluorescence. Using a cuvette-based time-resolved spectrofluorimeter employing one-photon excitation at 375 nm, we characterized the fluorescence intensity, lifetime, spectral response, anisotropy and time-resolved anisotropy of NADH in aqueous solution under varying microenvironmental conditions, namely temperature, pH, and binding to lactate dehydrogenase (LDH). Our results demonstrate how temperature, pH, and binding partners each impact the fluorescence signature of NADH and highlight the complexity of the fluorescence data when different parameters produce competing effects. We hope that the data presented in this study will provide a reference for potential sources of variation in experiments measuring NADH fluorescence.
Issue Date: 15-May-2021
Date of Acceptance: 25-Apr-2021
URI: http://hdl.handle.net/10044/1/89545
DOI: 10.1364/OSAC.423082
Publisher: OSA Publishing
Start Page: 1610
End Page: 1625
Journal / Book Title: Optical Society of America Continuum
Volume: 4
Issue: 5
Copyright Statement: © 2021. Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI.
Sponsor/Funder: Engineering & Physical Science Research Council (EPSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: EP/I02770X/1
BB/E000495/1
Keywords: Science & Technology
Physical Sciences
Optics
NICOTINAMIDE ADENINE-DINUCLEOTIDE
MULTIPHOTON EXCITATION
REDUCED NICOTINAMIDES
INTRACELLULAR NADH
DECAY KINETICS
LIFETIME
DYNAMICS
NAD(P)H
CELLS
STATE
Publication Status: Published
Online Publication Date: 2021-05-15
Appears in Collections:Physics
Department of Surgery and Cancer
Photonics
Faculty of Natural Sciences



This item is licensed under a Creative Commons License Creative Commons