A common property during tumor development and other diseases is altered energy metabolism, which could lead to a switch between oxidative phosphorylation (OXPHOS) and glycolysis. FLIM of metabolic coenzymes, as NAD(P)H and FAD, is now widely accepted to be the most reliable method to determine cell metabolism and different algorithms are actually proved to get reproducible results. However, it is questionable if redox state and cell metabolism correlate. For this new algorithms has to be developed to improve the diagnostic procedure. We will discuss the different approaches which were published so far and present new results in metabolic imaging taking into account the FLIM based NADH metabolic index as well as FLIRR based calculations including NADH, FAD and FMN. In addition, the phosphorescence lifetime of newly developed drugs will be used to demonstrate local oxygen changes. Simultaneous imaging of phosphorescence and fluorescence lifetime parameters enables complex analysis of bioenergetic alterations.
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