The bh DCS-120 Confocal Scanning FLIM System detects changes in the metabolic state of live cells. Information on the metabolic state is derived from the fluorescence decay functions of NAD(P)H and FAD. Two picosecond diode lasers, with wavelengths of 375 nm and 405 nm, are multiplexed to alternatingly excite NAD(P)H and FAD. One FLIM channel of the DCS-120 system detects in the emission band of NAD(P)H, the other in the emission band of FAD.
The FLIM data are processed by bh SPCImage data analysis software. For both channels, the data analysis delivers images of the amplitude-weighted lifetime, τm, the component lifetimes, τ1 and τ2, the amplitudes of the components, a1 and a2, and the amplitude ratio, a1/a2. Moreover, it delivers the fluorescence-lifetime redox ratio (FLIRR), a2(NADH)/a1(FAD). A shift from oxidative phosphorylation to glycolysis or back is revealed by changes in τm, a1 or a1/a2, and in the FLIRR.
For details please see: Metabolic Imaging with the DCS-120 Confocal FLIM System: Simultaneous FLIM of NAD(P)H and FAD
NAD(P)H a1 images for normal cells (left) and tumor cells (middle left). FAD a1 images for normal cells (middle right) and tumor cells (right).