Wide-field time-correlated single photon counting (TCSPC) microscopy with time resolution below the frame exposure time

Wide-field time-correlated single photon counting-based fluorescence lifetime imaging microscopy

“…This approach is similar to dual exposure techniques for velocity map imaging in mass spectroscopy [42], [43]. The measurement of the photon arrival time from the phosphor decay can improve the time resolution beyond the inverse frame rate of the camera, and the lower frame rate increases the number of recorded pixels, thus allowing bigger field of view [39], [44]. Phosphorescent sample decays as short as 500 ns have been measured with a P20 phosphor and 300 kHz frame rate [45], as shown in Fig.…”

Wide-field TCSPC: methods and applications

“…The measurement of the arrival time from the photon event phosphor decay can improve the time resolution beyond the inverse frame rate of the camera, and the slower frame rate used for this technique also increases the number of recorded pixels, thus allowing a bigger field of view. Lifetimes around 1 μs have been measured with several transition metal probes using a 54 kHz frame rate, including a ruthenium based oxygen sensor in living cells [187,188], and a lifetime of ∼500 ns has been measured with a 300 kHz camera frame rate [84].…”

Wide-field TCSPC-based fluorescence lifetime imaging (FLIM) microscopy

“…The measurement of the arrival time from the photon event phosphor decay can improve the time resolution beyond the inverse frame rate of the camera, and the lower frame rate increases the number of recorded pixels, thus allowing bigger field of view. 20,27 Phosphorescent sample decays as short as 500 ns have been measured with a P20 phosphor and 300 kHz frame rate, 21 but a combination of a faster phosphor and a faster frame rate, or special cameras such as timepix 28 or pimms, 29 could allow the measurement of even faster sample decays.…”