Time-resolved fluorescence

By imaging the fluorescence lifetime and/or time-resolved anisotropy via time-correlated single photon counting (TCSPC), detailed insights can be obtained about the fluorophore’s photophysical properties, which is important for quantitative and absolute interpretations of many fluorescence experiments. In our research we specialize in the development of multicolor TCSPC methods by using nanosecond alternating excitation (aka pulsed interleaved excitation, PIE) (Hendrix et al., 2013 and Hendrix et al., 2014).

To avoid pixel-based decay fitting, and complementary to whole-image FLIM analysis, we also specialize in advanced Phasor-FLIM analysis. This is a graphical way (via the phasor plot) to false-color intensity images with lifetimes, lifetime fractions or FRET efficiencies.

Furthermore, we combine time-resolved spectroscopy with imaging and fluctuation methods, which allows detailed investigations of complex biological systems.

We also specialize in steady state and time-resolved anisotropy, whether or not in combination with imaging. Our commercial confocal LSM880 is also equipped for polarized SHG.