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Fast In Vivo Imaging of SHG Nanoprobes with Multiphoton Light-Sheet Microscopy

Abstract : Two-photon light-sheet microscopy (2P-SPIM) provides a unique combination of advantages for fast and deep fluorescence imaging in live tissues. Detecting coherent signals such as second-harmonic generation (SHG) in 2P-SPIM in addition to fluorescence would open further imaging opportunities. However, light-sheet microscopy involves an orthogonal configuration of illumination and detection that questions the ability to detect coherent signals. Indeed, coherent scattering from micron-sized structures occurs predominantly along the illumination beam. By contrast, point-like sources such as SHG nanocrystals can efficiently scatter light in multiple directions and be detected using the orthogonal geometry of a light-sheet microscope. This study investigates the suitability of SHG light-sheet microscopy (SHG-SPIM) for fast imaging of SHG nanoprobes. Parameters that govern the detection efficiency of KTiOPO 4 and BaTiO 3 nanocrystals using SHG-SPIM are investigated theoretically and experimentally. The effects of incident polarization, detection numerical aperture, nanocrystal rotational motion, and second-order susceptibility tensor symmetries on the detectability of SHG nanoprobes in this specific geometry are clarified. Guidelines for optimizing SHG-SPIM imaging are established, enabling fast in vivo light-sheet imaging combining SHG and two-photon excited fluorescence. Finally, microangiography was achieved in live zebrafish embryos by SHG imaging at up to 180 frames per second and single-particle tracking of SHG nanoprobes in the blood flow.
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Contributor : Willy Supatto Connect in order to contact the contributor
Submitted on : Wednesday, November 25, 2020 - 11:30:13 AM
Last modification on : Wednesday, December 1, 2021 - 2:36:54 PM
Long-term archiving on: : Friday, February 26, 2021 - 6:51:17 PM


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Guy Malkinson, Pierre Mahou, Élodie Chaudan, Thierry Gacoin, Ali Sonay, et al.. Fast In Vivo Imaging of SHG Nanoprobes with Multiphoton Light-Sheet Microscopy. ACS photonics, American Chemical Society,, 2020, 7 (4), pp.1036-1049. ⟨10.1021/acsphotonics.9b01749⟩. ⟨hal-02958634⟩



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