The system presented here is a further development of the recently introduced spectro-temporal laser imaging by diffractive excitation (SLIDE) microscopy technique. To excite endogenous fluorescence, a new flexible and fibre-based laser source at 780 nm has been developed. The fiber-based FDML-MOPA was amplified to high peak and average powers using rare earth erbium fiber amplifiers. Broadband quasi-phase-matched frequency doubling using a fan-out PPLN crystal was then employed. The output is a 10 nm wide swept pulsed laser at 780 nm with a peak pulse power of 150 W, a pulse duration of 44 ps and a pulse repetition rate of 82 MHz (250 pulses at a sweep rate of 347 kHz). The sweep rate is converted into line scans by a diffraction grating and sent to a microscope for two-photon excitation of UV-excited dyes or endogenous autofluorescence. For detection, the signal is captured with a 4 GS/s high-speed digitisation card, resulting in 2kHz fluorescence lifetime imaging (FLIM) acquisition. In this paper we present the first images of 780nm SLIDE obtained at 2kHz frame rate. Through the additional use of a piezo objective scanner, we are able to perform 3D imaging at 20Hz volume rate. We have also used this novel system for high-speed LiDAR imaging at 2kHz using the recently introduced SLIDE-based time-stretch LiDAR approach.
The system presented here is an evolution of the recently introduced spectro-temporal laser imaging by diffractive excitation (SLIDE) (1) microscopy technique. In order to excite endogenous fluorescence, a new flexible and fiber-based laser source at 780 nm was developed. The fiber-based FDML-MOPA (2) was amplified to high peak and average powers by rare-earth Erbium fiber amplifiers. Afterwards, broadband quasi-phase-matched frequency-doubling using a Fan-out PPLN crystal was employed. The output is a 10 nm wide swept, pulsed laser around 780 nm with a pulse peak power of 150 W, 44 ps pulse duration and a pulse repetition rate of 82 MHz (250 pulses at 347 kHz sweep rate). The sweep rate is converted to line scans by a diffraction grating and sent to a microscope for two-photon excitation of UV-excited dyes or endogenous autofluorescence. For detection, the signal is captured with a 4 GS/s high speed digitization card leading to 2 kHz fluorescence lifetime imaging (FLIM) acquisition.
In this work, we present first images of 780 nm SLIDE obtained at 2 kHz frame-rate. Through the additional use of an piezo objective scanner, we are able to perform 3D imaging at 20 Hz volume rate. We have also used this novel system for high-speed LiDAR imaging at a frequency of 2 kHz, using the recently introduced SLIDE-based time-stretch LiDAR approach.
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