Theoretical and experimental investigation of the depth limit of three-photon microscopy

Najva Akbari; Chris Xu

doi:10.1117/12.2582771

5 March 2021 Theoretical and experimental investigation of the depth limit of three-photon microscopy

Najva Akbari, Chris Xu

Proceedings Volume 11648, Multiphoton Microscopy in the Biomedical Sciences XXI; 116481A (2021) https://doi.org/10.1117/12.2582771
Event: SPIE BiOS, 2021, Online Only

Abstract

Multiphoton microscopy (MPM) techniques provide unprecedented deep access into live scattering tissue. Three-photon microscopy (3PM) has the advantage of the highly confined excitation volume, and its excitation wavelength falls in the optimal wavelength window for deep imaging of common fluorophores in scattering media. These features enable deep high-resolution imaging in complex biological tissues. Here, we provide a theoretical prediction for the depth limit in 3PM and experimentally verify our results. We show both theoretically and experimentally that the depth limit of 3PM depends on the labeling density of the sample, and 3PM can penetrate beyond 10 effective attenuation lengths (EAL) with close to diffraction-limited spatial resolution in a tissue phantom.

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Citation Download Citation

Najva Akbari and Chris Xu "Theoretical and experimental investigation of the depth limit of three-photon microscopy", Proc. SPIE 11648, Multiphoton Microscopy in the Biomedical Sciences XXI, 116481A (5 March 2021); https://doi.org/10.1117/12.2582771

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