IN2SIGHT: a microstructured window for label-free non-linear imaging in vivo.

Giuseppe Chirico; Laura Sironi; Davide Panzeri; Claudio Conci; Donato Inverso; Emanuela Jacchetti; Rebeca Martínez Vázquez; Roberto Osellame; Maddalena Collini; Giulio Cerullo; Manuela Teresa Raimondi

doi:10.1117/12.3026420

20 June 2024 IN2SIGHT: a microstructured window for label-free non-linear imaging in vivo.

Giuseppe Chirico, Laura Sironi, Davide Panzeri, Claudio Conci, Donato Inverso, Emanuela Jacchetti, Rebeca Martínez Vázquez, Roberto Osellame, Maddalena Collini, Giulio Cerullo, Manuela Teresa Raimondi

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Proceedings Volume PC13006, Biomedical Spectroscopy, Microscopy, and Imaging III; PC1300616 (2024) https://doi.org/10.1117/12.3026420
Event: SPIE Photonics Europe, 2024, Strasbourg, France

Abstract

Tissue histopathology, reliant on costly and time-consuming hematoxylin and eosin (H&E) staining of thin tissue slices, faces limitations. Label-free non-linear optical microscopy in vivo presents a solution, allowing work on fresh samples. Implantable microstructures prove effective for systematic longitudinal in vivo studies of immunological responses to biomaterials using label-free non-linear optical microscopy. Employing two-photon laser polymerization, we implanted a matrix of 3D lattices in the chorioallantoic membrane of chicken embryos, establishing a 3D reference frame for cell counting. H&E analysis is compared to label-free in vivo non-linear excitation imaging for cell quantification and identifying granulocytes, collagen, and microvessels. Preliminary results in higher animal models demonstrate the transformative potential of this approach, offering an alternative to conventional histopathology for validating biomaterials in in vivo longitudinal studies.

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Giuseppe Chirico, Laura Sironi, Davide Panzeri, Claudio Conci, Donato Inverso, Emanuela Jacchetti, Rebeca Martínez Vázquez, Roberto Osellame, Maddalena Collini, Giulio Cerullo, and Manuela Teresa Raimondi "IN2SIGHT: a microstructured window for label-free non-linear imaging in vivo.", Proc. SPIE PC13006, Biomedical Spectroscopy, Microscopy, and Imaging III, PC1300616 (20 June 2024); https://doi.org/10.1117/12.3026420

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KEYWORDS

In vivo imaging

Tissues

Windows

Animal model studies

Microlens

Nonlinear optics

Histopathology

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