Advances in label-free multimodal multiphoton imaging of living tissues

Chiara Stringari

doi:10.1117/12.3025177

18 June 2024 Advances in label-free multimodal multiphoton imaging of living tissues

Chiara Stringari

Proceedings Volume 12996, Unconventional Optical Imaging IV; 129960E (2024) https://doi.org/10.1117/12.3025177
Event: SPIE Photonics Europe, 2024, Strasbourg, France

Abstract

Many current biomedical applications require the development of advanced non-linear optical microscopy methods to provide functional and non-invasive imaging of live tissues with sub-cellular resolution. In the first part of the talk I will present recent advances in label-free metabolic imaging of living tissues by two-photon fluorescence lifetime microscopy (2P-FLIM) of endogenous biomarkers. We implement simultaneous two-photon excitation of NAD(P)H and FAD by wavelength mixing to acquire 2P-FLIM data of the two biomarkers at the same time and perform efficient multiparametric metabolic imaging in dynamic biological system. We demonstrate how 2P-FLIM of the two metabolic coenzymes reveals the richness and complexity of several metabolic processes in intact tissues with minimal phototoxicity and can be implemented non-invasively for longitudinal studies in vitro and in vivo such as stem cell differentiation, T cell activation and embryo development. In the second part of the talk I will present recent advances in polarization resolved second harmonic generation (pSHG) and modelling from protein molecular structure. We establish a general multi-scale numerical framework relating the micrometer-scale SHG measurements at the optical wavelength to the atomic-scale and molecular structure of the proteins under study and their supramolecular arrangement.

Conference Presentation

Citation Download Citation

Chiara Stringari "Advances in label-free multimodal multiphoton imaging of living tissues", Proc. SPIE 12996, Unconventional Optical Imaging IV, 129960E (18 June 2024); https://doi.org/10.1117/12.3025177

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