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Metabolic Profiling of Live Cancer Tissues Using NAD(P)H Fluorescence Lifetime Imaging

  • Thomas S. BlackerEmail author
  • Michael D. E. Sewell
  • Gyorgy Szabadkai
  • Michael R. Duchen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1928)

Abstract

Altered metabolism is a hallmark of cancer, both resulting from and driving oncogenesis. The NAD and NADP redox couples play a key role in a large number of the metabolic pathways involved. In their reduced forms, NADH and NADPH, these molecules are intrinsically fluorescent. As the average time for fluorescence to be emitted following excitation by a laser pulse, the fluorescence lifetime, is exquisitely sensitive to changes in the local environment of the fluorophore, imaging the fluorescence lifetime of NADH and NADPH offers the potential for label-free monitoring of metabolic changes inside living tumors. Here, we describe the biological, photophysical, and methodological considerations required to establish fluorescence lifetime imaging (FLIM) of NAD(P)H as a routine method for profiling the metabolism of living cancer cells and tissues.

Key words

NADH NADPH Autofluorescence Fluorescence lifetime imaging Live-cell microscopy Cancer metabolism 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Thomas S. Blacker
    • 1
    • 2
    • 3
    Email author
  • Michael D. E. Sewell
    • 1
  • Gyorgy Szabadkai
    • 1
    • 2
    • 4
    • 5
  • Michael R. Duchen
    • 1
    • 2
  1. 1.Research Department of Cell & Developmental BiologyUniversity College LondonLondonUK
  2. 2.UCL Consortium for Mitochondrial ResearchUniversity College LondonLondonUK
  3. 3.Department of Physics & AstronomyUniversity College LondonLondonUK
  4. 4.The Francis Crick InstituteLondonUK
  5. 5.Department of Biomedical SciencesUniversity of PaduaPaduaItaly

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