Multiplexed fluorescence lifetime measurements by frequency-sweeping Fourier spectroscopy

We report simultaneous measurements of fluorescence lifetimes at multiple excitation wavelengths with a Fourier transform frequency domain fluorescence lifetime spectrometer. The spectrometer uses a Michelson interferometer with its differential optical path length scanning at a 22,000 Hz scan rate. The scan speed of the optical delay varies linearly during each scan and creates interference modulations that sweep from -150 to 150 MHz in 45.5 micros. The frequency-sweeping modulation allows nanosecond fluorescence lifetime measurements within 45.5 micros. Because the interference modulation frequency is wavelength dependent, under the Fourier multiplexing principle, the spectrometer can perform lifetime measurements on multiple excitation wavelengths simultaneously.