Cargando…
Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques
Metabolic FLIM (fluorescence lifetime imaging) is used to image bioenergetic status in cells and tissue. Whereas an attribution of the fluorescence lifetime of coenzymes as an indicator for cell metabolism is mainly accepted, it is debated whether this is valid for the redox state of cells. In this...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8199032/ https://www.ncbi.nlm.nih.gov/pubmed/34073057 http://dx.doi.org/10.3390/ijms22115952 |
_version_ | 1783707280413294592 |
---|---|
author | Kalinina, Sviatlana Freymueller, Christian Naskar, Nilanjon von Einem, Bjoern Reess, Kirsten Sroka, Ronald Rueck, Angelika |
author_facet | Kalinina, Sviatlana Freymueller, Christian Naskar, Nilanjon von Einem, Bjoern Reess, Kirsten Sroka, Ronald Rueck, Angelika |
author_sort | Kalinina, Sviatlana |
collection | PubMed |
description | Metabolic FLIM (fluorescence lifetime imaging) is used to image bioenergetic status in cells and tissue. Whereas an attribution of the fluorescence lifetime of coenzymes as an indicator for cell metabolism is mainly accepted, it is debated whether this is valid for the redox state of cells. In this regard, an innovative algorithm using the lifetime characteristics of nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD) to calculate the fluorescence lifetime induced redox ratio (FLIRR) has been reported so far. We extended the FLIRR approach and present new results, which includes FLIM data of the various enzymes, such as NAD(P)H, FAD, as well as flavin mononucleotide (FMN). Our algorithm uses a two-exponential fitting procedure for the NAD(P)H autofluorescence and a three-exponential fit of the flavin signal. By extending the FLIRR approach, we introduced FLIRR1 as protein-bound NAD(P)H related to protein-bound FAD, FLIRR2 as protein-bound NAD(P)H related to free (unbound) FAD and FLIRR3 as protein-bound NAD(P)H related to protein-bound FMN. We compared the significance of extended FLIRR to the metabolic index, defined as the ratio of protein-bound NAD(P)H to free NAD(P)H. The statistically significant difference for tumor and normal cells was found to be highest for FLIRR1. |
format | Online Article Text |
id | pubmed-8199032 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-81990322021-06-14 Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques Kalinina, Sviatlana Freymueller, Christian Naskar, Nilanjon von Einem, Bjoern Reess, Kirsten Sroka, Ronald Rueck, Angelika Int J Mol Sci Article Metabolic FLIM (fluorescence lifetime imaging) is used to image bioenergetic status in cells and tissue. Whereas an attribution of the fluorescence lifetime of coenzymes as an indicator for cell metabolism is mainly accepted, it is debated whether this is valid for the redox state of cells. In this regard, an innovative algorithm using the lifetime characteristics of nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD) to calculate the fluorescence lifetime induced redox ratio (FLIRR) has been reported so far. We extended the FLIRR approach and present new results, which includes FLIM data of the various enzymes, such as NAD(P)H, FAD, as well as flavin mononucleotide (FMN). Our algorithm uses a two-exponential fitting procedure for the NAD(P)H autofluorescence and a three-exponential fit of the flavin signal. By extending the FLIRR approach, we introduced FLIRR1 as protein-bound NAD(P)H related to protein-bound FAD, FLIRR2 as protein-bound NAD(P)H related to free (unbound) FAD and FLIRR3 as protein-bound NAD(P)H related to protein-bound FMN. We compared the significance of extended FLIRR to the metabolic index, defined as the ratio of protein-bound NAD(P)H to free NAD(P)H. The statistically significant difference for tumor and normal cells was found to be highest for FLIRR1. MDPI 2021-05-31 /pmc/articles/PMC8199032/ /pubmed/34073057 http://dx.doi.org/10.3390/ijms22115952 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Kalinina, Sviatlana Freymueller, Christian Naskar, Nilanjon von Einem, Bjoern Reess, Kirsten Sroka, Ronald Rueck, Angelika Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques |
title | Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques |
title_full | Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques |
title_fullStr | Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques |
title_full_unstemmed | Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques |
title_short | Bioenergetic Alterations of Metabolic Redox Coenzymes as NADH, FAD and FMN by Means of Fluorescence Lifetime Imaging Techniques |
title_sort | bioenergetic alterations of metabolic redox coenzymes as nadh, fad and fmn by means of fluorescence lifetime imaging techniques |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8199032/ https://www.ncbi.nlm.nih.gov/pubmed/34073057 http://dx.doi.org/10.3390/ijms22115952 |
work_keys_str_mv | AT kalininasviatlana bioenergeticalterationsofmetabolicredoxcoenzymesasnadhfadandfmnbymeansoffluorescencelifetimeimagingtechniques AT freymuellerchristian bioenergeticalterationsofmetabolicredoxcoenzymesasnadhfadandfmnbymeansoffluorescencelifetimeimagingtechniques AT naskarnilanjon bioenergeticalterationsofmetabolicredoxcoenzymesasnadhfadandfmnbymeansoffluorescencelifetimeimagingtechniques AT voneinembjoern bioenergeticalterationsofmetabolicredoxcoenzymesasnadhfadandfmnbymeansoffluorescencelifetimeimagingtechniques AT reesskirsten bioenergeticalterationsofmetabolicredoxcoenzymesasnadhfadandfmnbymeansoffluorescencelifetimeimagingtechniques AT srokaronald bioenergeticalterationsofmetabolicredoxcoenzymesasnadhfadandfmnbymeansoffluorescencelifetimeimagingtechniques AT rueckangelika bioenergeticalterationsofmetabolicredoxcoenzymesasnadhfadandfmnbymeansoffluorescencelifetimeimagingtechniques |