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Application of light‐emitting diodes (LEDs) in frequency‐domain fluorescence lifetime imaging microscopy (FLIM) has been limited by the trade‐off between modulation frequency and illumination intensity of LEDs, which affects the signal‐to‐noise ratio in fluorescence lifetime measurements. To increase modulation frequency without sacrificing output power of LEDs, we propose to use LEDs with multiple...
A spectrograph with continuous wavelength resolution has been integrated into a frequency‐domain fluorescence lifetime‐resolved imaging microscope (FLIM). The spectral information assists in the separation of multiple lifetime components, and helps resolve signal cross‐talking that can interfere with an accurate analysis of multiple lifetime processes. This extends the number of different dyes that...
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