Time-Resolved Spectrometry of Mitochondrial NAD(P)H Fluorescence and Its Applications for Evaluating the Oxidative State in Living Cells

  • Julia Horilova
  • Hauke Studier
  • Zuzana Nadova
  • Pavol Miskovsky
  • Dusan ChorvatJr.
  • Alzbeta Marcek ChorvatovaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1264)


Time-resolved fluorescence spectrometry is a highly valuable technological tool to detect and characterize mitochondrial metabolic oxidative changes by means of endogenous fluorescence (Chorvat and Chorvatova, Laser Phys Lett 6: 175–193, 2009). Here, we describe the detection and measurement of endogenous mitochondrial NAD(P)H (nicotinamide adenine dinucleotide (phosphate)) fluorescence directly in living cultured cells using fluorescence lifetime spectrometry imaging after excitation with 405 nm picosecond (ps) laser. Time-correlated single photon counting (TCSPC) method is employed.

Key words

Mitochondrial oxidative state Endogenous NAD(P)H fluorescence FLIM Time-resolved spectrometry Energy metabolism 



Authors acknowledge support from Integrated Initiative of European Laser Research Infrastructures LASERLAB-EUROPE III (grant agreement no 284464, EC’s Seventh Framework Programme), EC’s Seventh Framework Programme CELIM 316310 project, the Research Grant Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic VEGA No 1/0296/11, and the Slovak Research and Development Agency under the contract APVV-0242-11.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Julia Horilova
    • 1
    • 2
  • Hauke Studier
    • 3
  • Zuzana Nadova
    • 2
    • 4
  • Pavol Miskovsky
    • 2
    • 4
  • Dusan ChorvatJr.
    • 1
  • Alzbeta Marcek Chorvatova
    • 5
    • 1
    Email author
  1. 1.Department of BiophotonicsInternational Laser CenterBratislavaSlovakia
  2. 2.Department of Biophysics, Faculty of SciencePavol Jozef Safarik UniversityKosiceSlovakia
  3. 3.Becker & Hickl GmbHBerlinGermany
  4. 4.Centre for Interdisciplinary Biosciences (CIB), Faculty of SciencePavol Jozef Safarik UniversityKosiceSlovakia
  5. 5.Department of Biotechnology, Faculty of Natural SciencesUniversity of Ss. Cyril and MethodiusTrnavaSlovakia

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