Methods of time resolved spectroscopy are applied in biomedicine during diagnosis of various malignant and nonmalignant diseases. Fluorescence lifetime measurement provides monitoring of intracellular metabolic enzymes like NADH and FAD, controlling the balance between oxidative phosphorylation and glycolysis. The simultaneous monitoring of the different metabolic parameters is still a challenge.

Here we present a setup, which is based on two-photon microscopy and multi-dimensional time-correlated single-photoncounting (TCSPC). The presented method allows consecutive NADH and FAD imaging with high specificity and minimal side effects. In addition the setup is useful for simultaneous monitoring of NADH FLIM and intracellular oxygen partial pressure (pO₂). With the two-channel FLIM/PLIM system we could show that pO₂ is investigated simultaneously with different metabolic parameters in living cells.

Date Published: 5 March 2015
PDF: 5 pages
Proc. SPIE 9329, Multiphoton Microscopy in the Biomedical Sciences XV, 93290C (5 March 2015); doi: 10.1117/12.2079166

Published in SPIE Proceedings Vol. 9329:
Multiphoton Microscopy in the Biomedical Sciences XV
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)