Probing of cardiomyocyte metabolism by spectrally resolved lifetime detection of NAD(P)H fluorescence

Aneba, Swida; Cheng, Ying; Mateášik, Anton; Comte, Blandine; Chorvát, D.; Chorvatova, Alzbeta

doi:10.1109/cic.2007.4745493

Cited by 2 publications

(1 citation statement)

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“…In living cardiac cells, we have estimated spectra, fluorescence lifetimes and pre-exponential factors of the flavin AF decays in steady-state conditions [19], as well as during field-stimulated contraction of rat cardiomyocytes [142]. We have also determined spectra and lifetime values of NAD(P)H in rat [143,144], as well as human [144] cardiac myocytes. In addition, we have developed analytical methods for the separation of individual components in spectrally-resolved confocal images [27] and time-resolved signals from cell AF [29].…”

Section: Identification Of Oxidative Metabolic Changes In Metabolical...mentioning

confidence: 99%

Multi-wavelength fluorescence lifetime spectroscopy: a new approach to the study of endogenous fluorescence in living cells and tissues

2009

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The study of biological systems in their real environmental conditions is crucial to decipher the true image of structures and processes underlying their functionality. In this regard, development of non‐invasive optical techniques that do not require labelling, such as the investigation of tissue endogenous fluorescence, is particularly important and, as reflected in the increasing number of contributions published recently on this subject, was recognized by many leading groups. Multi‐spectral and lifetime detection of fluorescence provides an effective experimental tool to discriminate between multiple naturally‐occurring fluorophores in living tissues. At the same time, however, data analysis allowing us to understand the spectral, temporal and spatial information gathered, describing individual molecules involved in the autofluorescence of intact biological systems, represents a tough scientific challenge that has not yet been fully resolved. In this review, we discuss the latest advances in technologies that record and assess spectrally‐resolved fluorescence lifetime data as well as their biological and clinical applications. We show how these methods provide efficient sensing of molecules correlated with changes in the mitochondrial metabolic redox state in pathological conditions and/or of cell ultrastructures in diseased tissue, based on the presence of oxidation/reductionsensitive fluorophores and/or cell‐specific chromophores. Future directions are also outlined. (© 2009 by Astro Ltd., Published exclusively by WILEY‐VCH Verlag GmbH & Co. KGaA)

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Section: Identification Of Oxidative Metabolic Changes In Metabolical...mentioning

confidence: 99%

Multi-wavelength fluorescence lifetime spectroscopy: a new approach to the study of endogenous fluorescence in living cells and tissues

2009

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Time-resolved fluorescence spectroscopy investigation of the effect of 4-hydroxynonenal on endogenous NAD(P)H in living cardiac myocytes

et al. 2013

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Lipid peroxidation is a major biochemical consequence of the oxidative deterioration of polyunsaturated lipids in cell membranes and causes damage to membrane integrity and loss of protein function. 4-hydroxy-2-nonenal (HNE), one of the most reactive products of n-6 polyunsaturated fatty acid peroxidation of membrane phospholipids, has been shown to be capable of affecting both nicotinamide adenine dinucleotide (phosphate) reduced [NAD(P)H] as well as NADH production. However, the understanding of its effects in living cardiac cells is still lacking. Our goal was to therefore investigate HNE effects on NAD(P)H noninvasively in living cardiomyocytes. Spectrally resolved lifetime detection of endogenous fluorescence, an innovative noninvasive technique, was employed. Individual fluorescence components were resolved by spectral linear unmixing approach. Gathered results revealed that HNE reduced the amplitude of both resolved NAD(P)H components in a concentration-dependent manner. In addition, HNE increased flavoprotein fluorescence and responsiveness of the NAD(P)H component ratio to glutathione reductase (GR) inhibitor. HNE also increased the percentage of oxidized nucleotides and decreased maximal NADH production. Presented data indicate that HNE provoked an important cell oxidation by acting on NAD(P)H regulating systems in cardiomyocytes. Understanding the precise role of oxidative processes and their products in living cells is crucial for finding new noninvasive tools for biomedical diagnostics of pathophysiological states.

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Probing of cardiomyocyte metabolism by spectrally resolved lifetime detection of NAD(P)H fluorescence

Cited by 2 publications

References 4 publications

Multi-wavelength fluorescence lifetime spectroscopy: a new approach to the study of endogenous fluorescence in living cells and tissues

Multi-wavelength fluorescence lifetime spectroscopy: a new approach to the study of endogenous fluorescence in living cells and tissues

Time-resolved fluorescence spectroscopy investigation of the effect of 4-hydroxynonenal on endogenous NAD(P)H in living cardiac myocytes

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