Non-linear excitation microscopy offers several advantages for in-vivo imaging compared to conventional confocal techniques. However, tissue penetration can still be an issue due to scattering and spherical aberrations induced on focused beams by the tissue. The use of low numerical aperture objectives to pass through the outer layers of the skin, together with high dioptric power microlenses implanted in-vivo close to the observation volume, can be beneficial to the reduction of optical aberrations. Here, Fibroblast cell culture plano-convex microlenses to be used for non-linear imaging of biological tissue are developed and tested. The microlenses can be used as single lenses or multiplexed in an array. A thorough test of the lenses wavefront is reported together with the modulation transfer function and wavefront profile. Magnified fluorescence images can be retrieved through the microlenses coupled to commercial confocal and two-photon excitation scanning microscopes. The signal-to-noise ratio of the images is not substantially affected by the use of the microlenses and the magnification can be adjusted by changing the relative position of the microlens array to the microscope objective and the immersion medium. These results are opening the way to the application of implanted micro-optics for optical in-vivo inspection of biological processes.

Marini, M., Nardini, A., Martínez Vázquez, R., Conci, C., Bouzin, M., Collini, M., et al. (2023). Microlenses Fabricated by Two‐Photon Laser Polymerization for Cell Imaging with Non‐Linear Excitation Microscopy. ADVANCED FUNCTIONAL MATERIALS, 33(39 (September 26, 2023)) [10.1002/adfm.202213926].

Microlenses Fabricated by Two‐Photon Laser Polymerization for Cell Imaging with Non‐Linear Excitation Microscopy

Marini, M.;Bouzin, M.;Collini, M.;Chirico, G.
2023

Abstract

Non-linear excitation microscopy offers several advantages for in-vivo imaging compared to conventional confocal techniques. However, tissue penetration can still be an issue due to scattering and spherical aberrations induced on focused beams by the tissue. The use of low numerical aperture objectives to pass through the outer layers of the skin, together with high dioptric power microlenses implanted in-vivo close to the observation volume, can be beneficial to the reduction of optical aberrations. Here, Fibroblast cell culture plano-convex microlenses to be used for non-linear imaging of biological tissue are developed and tested. The microlenses can be used as single lenses or multiplexed in an array. A thorough test of the lenses wavefront is reported together with the modulation transfer function and wavefront profile. Magnified fluorescence images can be retrieved through the microlenses coupled to commercial confocal and two-photon excitation scanning microscopes. The signal-to-noise ratio of the images is not substantially affected by the use of the microlenses and the magnification can be adjusted by changing the relative position of the microlens array to the microscope objective and the immersion medium. These results are opening the way to the application of implanted micro-optics for optical in-vivo inspection of biological processes.
Articolo in rivista - Articolo scientifico
3D micro scaffolds; confocal microscopy; microlenses; SZ2080; two-photon imaging; two-photon polymerization;
English
15-mar-2023
2023
33
39 (September 26, 2023)
2213926
none
Marini, M., Nardini, A., Martínez Vázquez, R., Conci, C., Bouzin, M., Collini, M., et al. (2023). Microlenses Fabricated by Two‐Photon Laser Polymerization for Cell Imaging with Non‐Linear Excitation Microscopy. ADVANCED FUNCTIONAL MATERIALS, 33(39 (September 26, 2023)) [10.1002/adfm.202213926].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/405555
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