A bimodal fluorescence and phase microscope for time-lapse imaging of 3D multi-scattering objects

Chiara Paviolo; Sylvia Desissaire; Edgar del Llano; Marie Fackeure; Jeremy Neri; Cécile Fiche; Corinne Loeuillet; Pierre F. Ray; Christophe Arnoult; Kiran Padmanabhan; Lionel Hervé

doi:10.1117/12.3017387

18 June 2024 A bimodal fluorescence and phase microscope for time-lapse imaging of 3D multi-scattering objects

Chiara Paviolo, Sylvia Desissaire, Edgar del Llano, Marie Fackeure, Jeremy Neri, Cécile Fiche, Corinne Loeuillet, Pierre F. Ray, Christophe Arnoult, Kiran Padmanabhan, Lionel Hervé

Author Affiliations +

Proceedings Volume PC12996, Unconventional Optical Imaging IV; PC129960P (2024) https://doi.org/10.1117/12.3017387
Event: SPIE Photonics Europe, 2024, Strasbourg, France

Abstract

We present a diffraction-phase and fluorescence 3D microscope as novel bimodal imaging technique, which provides simultaneous phase and multi-color epi-fluorescence acquisitions of living multicellular samples. The instrument consists of an LED-array to acquire intensity images at different illumination angles and an epifluorescence setup for fluorescence excitation. The 3D sample’s optical properties are reconstructed using the beam propagation method embedded inside a deep learning framework. To obtain the fluorescence reconstructions, we developed a novel incoherent model that takes into account the heterogeneous refractive indexes of the scattering sample. We validated the technique on long-term acquisitions of mouse embryos and 3D liver organoids under physiological conditions.

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Chiara Paviolo, Sylvia Desissaire, Edgar del Llano, Marie Fackeure, Jeremy Neri, Cécile Fiche, Corinne Loeuillet, Pierre F. Ray, Christophe Arnoult, Kiran Padmanabhan, and Lionel Hervé "A bimodal fluorescence and phase microscope for time-lapse imaging of 3D multi-scattering objects", Proc. SPIE PC12996, Unconventional Optical Imaging IV, PC129960P (18 June 2024); https://doi.org/10.1117/12.3017387

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