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Modeling nonlinear microscopy near index-mismatched interfaces

Abstract : Nonlinear microscopy is widely used to characterize thick, optically heterogeneous biological samples. While quantitative image analysis requires accurately describing the contrast mechanisms at play, the majority of established numerical models neglect the influence of field distortion caused by sample heterogeneity near focus. In this work, we show experimentally and numerically that finite-difference time-domain (FDTD) methods are applicable to model focused fields interactions in the presence of heterogeneities, typical of nonlinear microscopy. We analyze the ubiquitous geometry of a vertical interface between index-mismatched media (water, glass, and lipids) and consider the cases of two-photon-excited fluorescence (2PEF), third-harmonic generation (THG) and polarized THG contrasts. We show that FDTD simulations can accurately reproduce experimental images obtained on model samples and in live adult zebrafish, in contrast with previous models neglecting field distortions caused by index mismatch at the micrometer scale. Accounting for these effects appears to be particularly critical when interpreting coherent and polarization-resolved microscopy data.
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Contributor : Emmanuel Beaurepaire Connect in order to contact the contributor
Submitted on : Friday, July 16, 2021 - 10:17:50 AM
Last modification on : Saturday, November 27, 2021 - 5:41:29 PM
Long-term archiving on: : Sunday, October 17, 2021 - 6:27:28 PM


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Josephine Morizet, Giovanni Sartorello, Nicolas Dray, Chiara Stringari, Emmanuel Beaurepaire, et al.. Modeling nonlinear microscopy near index-mismatched interfaces. Optica, Optical Society of America - OSA Publishing, 2021, 8 (7), pp.944. ⟨10.1364/OPTICA.421257⟩. ⟨hal-03287976⟩



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