Kilohertz two-photon SLIDE microscopy using a newly developed 780 nm 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:10.1117/12.2650540

16 March 2023 Kilohertz two-photon SLIDE microscopy using a newly developed 780 nm 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 12390, High-Speed Biomedical Imaging and Spectroscopy VIII; 1239007 (2023) https://doi.org/10.1117/12.2650540
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

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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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 780 nm excitation laser", Proc. SPIE 12390, High-Speed Biomedical Imaging and Spectroscopy VIII, 1239007 (16 March 2023); https://doi.org/10.1117/12.2650540

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