Kilohertz two-photon SLIDE microscopy using a newly developed 780nm excitation laser

Tonio F. Kutscher; Christian Stock; Florian Sommer; Jonas Jurkevicius; Stefan Meyer; Anton Gruber; Annika Hunold; Moritz Wiggert; Christina Leonhardt; Philipp Lamminger; Sebastian Karpf

doi:10.1117/12.2670904

9 August 2023 Kilohertz two-photon SLIDE microscopy using a newly developed 780nm excitation laser

Tonio F. Kutscher, Christian Stock, Florian Sommer, Jonas Jurkevicius, Stefan Meyer, Anton Gruber, Annika Hunold, Moritz Wiggert, Christina Leonhardt, Philipp Lamminger, Sebastian Karpf

Author Affiliations +

Proceedings Volume 12630, Advances in Microscopic Imaging IV; 126300B (2023) https://doi.org/10.1117/12.2670904
Event: European Conferences on Biomedical Optics, 2023, Munich, Germany

Abstract

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.

Citation Download Citation

Tonio F. Kutscher, Christian Stock, Florian Sommer, Jonas Jurkevicius, Stefan Meyer, Anton Gruber, Annika Hunold, Moritz Wiggert, Christina Leonhardt, Philipp Lamminger, and Sebastian Karpf "Kilohertz two-photon SLIDE microscopy using a newly developed 780nm excitation laser", Proc. SPIE 12630, Advances in Microscopic Imaging IV, 126300B (9 August 2023); https://doi.org/10.1117/12.2670904

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