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FLIM and PLIM in biomedical research – An innovative way to combine autofluorescence and oxygen measurements

  • Sviatlana Kalinina EMAIL logo and Angelika Rück
Published/Copyright: October 20, 2016
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Photonics & Lasers in Medicine
From the journal Photonics & Lasers in Medicine Volume 5 Issue 4

Abstract:

Fluorescence lifetime imaging microcopy (FLIM) is successfully used to image the intracellular fluorescent coenzymes NAD(P)H and FAD+. The redox state of these coenzymes is a parameter which helps to reveal the metabolic status of living cells and tissues. However, metabolic reactions are strongly dependent on the intracellular oxygen level. One promising optical method to monitor oxygen in biomedical samples is phosphorescence lifetime imaging microscopy (PLIM). PLIM is based on oxygen-dependent quenching of the phosphorescence of so-called “phosphors”. In this way, PLIM enables measurement of the oxygen partial pressure (pO2) within living cells. This review describes the FLIM and PLIM approaches used in biomedical research, drawing particular attention to the techniques of simultaneous FLIM and PLIM, which provide correlative imaging of both the fluorescence lifetime of metabolic coenzymes and pO2-sensitive phosphorescence lifetime.

Zusammenfassung:

Für die intrazelluläre Detektion der Koenzyme NAD(P)H und FAD+ eignen sich besonders gut zeitaufgelöste Fluoreszenzverfahren, wie die Fluoreszenzlebenszeit-Mikroskopie (FLIM), mit deren Hilfe der Redoxzustand der Enzyme bestimmt werden kann. Dieser ist vom metabolischen Zustand der Zellen und vom Gewebe abhängig. Darüber hinaus ist der Zellmetabolismus auch vom intrazellulären Sauerstoffgehalt abhängig. Ein vielversprechendes optisches Verfahren, um die Sauerstoffkonzentration in biomedizinischen Proben zu überwachen, ist die Phosphoreszenzlebenszeit-Mikroskopie (PLIM). Die von der Sauerstoffkonzentration abhängige Phosphoreszenzabklingzeit einiger Verbindungen, sogenannter Phosphore, kann genutzt werden, um den Sauerstoffpartialdruck (pO2) im physiologischen Bereich zu messen. Der vorliegende Übersichtsartikel beschreibt FLIM- und PLIM-Ansätze in der biomedizinischen Forschung mit besonderem Fokus auf simultane FLIM- und PLIM-Techniken, die eine korrelative Bildgebung sowohl der Fluoreszenzlebenszeit metabolischer Enzyme, als auch der pO2-empfindlichen Phosphoreszenz erlauben.

Keywords: NAD(P)H; FLIM; PLIM; autofluorescence; pO2; cell metabolism; oxidative phosphorylation; glycolysis
Schlüsselwörter:: NAD(P)H; FLIM; PLIM; Autofluoreszenz; pO2; Zellmetabolismus; oxidative Phosphorylierung; Glykolyse

Acknowledgments

We would like to express our sincere thanks to W. Becker, P. Schäfer, and J. Breymayer for fruitful discussions in the past.

  1. Conflict of interest statement: The authors state no conflict of interest. All authors have read the journal’s Publication Ethics and Publication Malpractice Statement available at the journal’s website and hereby confirm that they comply with all its parts applicable to the present scientific work.

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Received: 2016-7-18
Revised: 2016-9-12
Accepted: 2016-9-20
Published Online: 2016-10-20
Published in Print: 2016-11-1

©2016 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Lasers, applications and technologies
  4. Editors’ notes
  5. Reviewer acknowledgment
  6. Photonics & Lasers in Medicine – Dissolved in diversity
  7. Announcement
  8. Pater Leander Fischer Prize 2016
  9. Magazine section
  10. Snapshots
  11. Review
  12. FLIM and PLIM in biomedical research – An innovative way to combine autofluorescence and oxygen measurements
  13. Congress report
  14. International Conference on Lasers, Applications and Technologies (LAT2016)
  15. Congress announcements
  16. Congresses 2016/2017
  17. Contents of the Volume
  18. Contents of the Volume
https://doi.org/10.1515/plm-2016-0026
Keywords for this article
NAD(P)H; FLIM; PLIM; autofluorescence; pO2; cell metabolism; oxidative phosphorylation; glycolysis

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Lasers, applications and technologies
  4. Editors’ notes
  5. Reviewer acknowledgment
  6. Photonics & Lasers in Medicine – Dissolved in diversity
  7. Announcement
  8. Pater Leander Fischer Prize 2016
  9. Magazine section
  10. Snapshots
  11. Review
  12. FLIM and PLIM in biomedical research – An innovative way to combine autofluorescence and oxygen measurements
  13. Congress report
  14. International Conference on Lasers, Applications and Technologies (LAT2016)
  15. Congress announcements
  16. Congresses 2016/2017
  17. Contents of the Volume
  18. Contents of the Volume
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