Fluorescence lifetime imaging for the two-photon microscope: time-domain and frequency-domain methods.
- Author(s): Gratton, Enrico
- Breusegem, Sophie
- Sutin, Jason
- Ruan, Qiaoqiao
- Barry, Nicholas
- et al.
Published Web Locationhttps://doi.org/10.1117/1.1586704
Fluorescence lifetime images are obtained with the laser scanning microscope using two methods: the time-correlated single-photon counting method and the frequency-domain method. In the same microscope system, we implement both methods. We perform a comparison of the performance of the two approaches in terms of signal-to-noise ratio (SNR) and the speed of data acquisition. While in our practical implementation the time-correlated single-photon counting technique provides a better SNR for low-intensity images, the frequency-domain method is faster and provides less distortion for bright samples.