Cell metabolism, FLIM and PLIM and applications

摘要

Methods of time resolved spectroscopy are applied in biomedicine during diagnosis of various malignant and non-malignant 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-photon-counting (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 (pO_2). With the two-channel FLIM/PLIM system we could show that pO_2 is investigated simultaneously with different metabolic parameters in living cells.