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Metabolic Imaging Using Two-Photon Excited NADH Intensity and Fluorescence Lifetime Imaging

Published online by Cambridge University Press:  26 July 2012

Jorge Vergen ,
Clifford Hecht ,
Lyandysha V. Zholudeva ,
Meg M. Marquardt ,
Richard Hallworth  and
Michael G. Nichols
Jorge Vergen
Affiliation:
Department of Physics, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA
Clifford Hecht
Affiliation:
Department of Physics, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA
Lyandysha V. Zholudeva
Affiliation:
Department of Physics, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA
Meg M. Marquardt
Affiliation:
Department of Physics, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA
Richard Hallworth
Affiliation:
Department of Biomedical Sciences, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA
Michael G. Nichols*
Affiliation:
Department of Physics, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA
*
Corresponding author. E-mail: [email protected]
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Abstract

Metabolism and mitochondrial dysfunction are known to be involved in many different disease states. We have employed two-photon fluorescence imaging of intrinsic mitochondrial reduced nicotinamide adenine dinucleotide (NADH) to quantify the metabolic state of several cultured cell lines, multicell tumor spheroids, and the intact mouse organ of Corti. Historically, fluorescence intensity has commonly been used as an indicator of the NADH concentration in cells and tissues. More recently, fluorescence lifetime imaging has revealed that changes in metabolism produce not only changes in fluorescence intensity, but also significant changes in the lifetimes and concentrations of free and enzyme-bound pools of NADH. Since NADH binding changes with metabolic state, this approach presents a new opportunity to track the cellular metabolic state.

Keywords

fluorescenceNADHmultiphoton microscopylifetime imaging
Type
Special Section: Seventh Omaha Imaging Symposium
Information
Microscopy and Microanalysis , Volume 18 , Issue 4 , August 2012 , pp. 761 - 770
Copyright
Copyright © Microscopy Society of America 2012

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References

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