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Oxygen sensing PLIM together with FLIM of intrinsic cellular fluorophores for metabolic mapping
Author(s): Sviatlana Kalinina; Patrick Schaefer; Jasmin Breymayer; Dominik Bisinger; Sabyasachi Chakrabortty; Angelika Rueck
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Paper Abstract

Otical imaging techniques based on time correlated single photon counting (TCSPC) has found wide applications in medicine and biology. Non-invasive and information-rich fluorescence lifetime imaging microscopy (FLIM) is successfully used for monitoring fluorescent intrinsic metabolic coenzymes as NAD(P)H (nicotinamide adenine dinucleotide (phosphate)) and FAD+ (flavin adenine dinucleotide) in living cells and tissues. The ratio between proteinbound and free coenzymes gives an information about the balance between oxidative phosphorylation and glycolysis in the cells. The changes of the ratio reflects major cellular disorders. A correlation exists between metabolic activity, redox ratio and fluorescence lifetime during stem cell differentiation, neurodegenerative diseases, and carcinogenesis.

A multichannel FLIM detection system was designed for monitoring the redox state of NAD(P)H and FAD+ and other intrinsic fluorophores as protoporphyrin IX. In addition, the unique upgrade is useful to perform FLIM and PLIM (phosphorescence lifetime imaging microscopy) simultaneously. PLIM is a promising method to investigate oxygen sensing in biomedical samples. In detail, the oxygen-dependent quenching of phosphorescence of some compounds as transition metal complexes enables measuring of oxygen partial pressure (pO2). Using a two-channel FLIM/PLIM system we monitored intrinsic pO2 by PLIM simultaneously with NAD(P)H by FLIM providing complex metabolic and redox imaging of living cells.

Physico-chemical properties of oxygen sensitive probes define certain parameters including their localisation. We present results of some ruthenium based complexes including those specifically bound to mitochondria.

Paper Details

Date Published: 20 February 2018
PDF: 6 pages
Proc. SPIE 10497, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVI, 104970F (20 February 2018); doi: 10.1117/12.2287429
Show Author Affiliations
Sviatlana Kalinina, Univ. Ulm (Germany)
Patrick Schaefer, Univ. Ulm (Germany)
Jasmin Breymayer, Univ. Ulm (Germany)
Dominik Bisinger, Univ. Ulm (Germany)
Sabyasachi Chakrabortty, Max-Planck-Institut für Polymerforschung (Germany)
Angelika Rueck, Univ. Ulm (Germany)


Published in SPIE Proceedings Vol. 10497:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XVI
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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