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Proceedings Paper

Phasor fluorescence lifetime microscopy of NADH to analyze metabolic activity of adipocytes (Conference Presentation)
Author(s): Michael Evers; Nunciada Salma; Sam Osseiran; Malte J. Casper; Reginald Birngruber; Conor L. Evans; Dieter Manstein
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Paper Abstract

The evaluation of complex metabolic changes of individual live cells and heterogeneous cell cultures is not feasible using traditional methods due to their destructive behavior and lack of spatial information. Two-photon excited fluorescence of intrinsic fluorophores such as nicotinamide adenine dinucleotide (NADH) facilitate a label-free and non-destructive evaluation of metabolic activity. This study explores the phasor approach in combination with two-photon fluorescence lifetime imaging microscopy (FLIM) as a potential method to evaluate pharmacologically induced metabolic changes that occur during the browning of adipocytes. The possibility of browning of white adipose cells is a desirable prospect for the treatment of obesity and related disorders. Here, we compared the results obtained by Fourier-based phasor analysis with the traditional exponential FLIM analysis as well as results of an extracellular flux analyzer. The alteration of glycolytic function and oxidative phosphorylation after treatment with pharmacological reagents significantly shift the contribution of each of the fluorescence lifetime components to the total fluorescence intensity. Further, we showed that the ratios of the lifetime components obtained by the phasor approach reflect the shift in mitochondrial and cytosolic NADH concentration. The phasor analysis agrees with traditional assessments, such as the optical free-to-bound NADH ratio as well as the oxygen consumption rate and extracellular acidification rate as determined by the extracellular flux analyzer. Our results support the concept that non-invasive sensing of fat metabolism and browning of fat may be possible by analyzing the fluorescence lifetime of NADH using the phasor approach.

Paper Details

Date Published: 14 March 2018
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Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII, 1049808 (14 March 2018); doi: 10.1117/12.2289294
Show Author Affiliations
Michael Evers, Massachusetts General Hospital, Harvard Univ. (United States)
Nunciada Salma, Massachusetts General Hospital, Harvard Univ. (United States)
Sam Osseiran, Massachusetts General Hospital, Harvard Univ. (United States)
Malte J. Casper, Massachusetts General Hospital, Harvard Univ. (United States)
Reginald Birngruber, Institut für Biomedizinische Optik, Univ. of Luebeck (Germany)
Conor L. Evans, Wellman Ctr. for Photomedicine, Massachusetts General Hospital (United States)
Dieter Manstein, Massachusetts General Hospital, Harvard Univ. (United States)


Published in SPIE Proceedings Vol. 10498:
Multiphoton Microscopy in the Biomedical Sciences XVIII
Ammasi Periasamy; Peter T. C. So; Karsten König; Xiaoliang S. Xie, Editor(s)

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