Polarimetric third-harmonic generation microscopy for histopathology

Mykolas Maciulis; Viktoras Mazeika; Vilte G. Samsone; Lukas Kontenis; Edvardas Zurauskas; Vitalijus Karabanovas; Virginijus Barzda

doi:10.1117/12.3001641

13 March 2024 Polarimetric third-harmonic generation microscopy for histopathology

Mykolas Maciulis, Viktoras Mazeika, Vilte G. Samsone, Lukas Kontenis, Edvardas Zurauskas, Vitalijus Karabanovas, Virginijus Barzda

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Proceedings Volume PC12846, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XXII; PC128460D (2024) https://doi.org/10.1117/12.3001641
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

In this work, multimodal two-photon excitation fluorescence (TPEF) and third-harmonic generation (THG) microscopy is used together with second-harmonic generation (SHG) to image gold standard hematoxylin and eosin histology slides. The nonlinear responses were investigated for hematoxylin and eosin stains. It is shown that the extracellular matrix (ECM) can be visualized using THG image contrast. THG signal is typically associated with interfaces and nanostructures where abrupt changes in the refractive index takes place, e.g. at blood vessel walls as well as the dense nuclei. Polarimetric THG provides a new method for ultrastructural ECM visualization. The ability of THG microscopy to visualize simultaneously nuclei and ECM is exemplified by imaging clinical tissue samples. THG turned out to be an excellent counterstaining contrast to SHG that reveals ECM proteins, cells and cell nuclei, which can be utilized for studying interactions between cells and ECM. Multiphoton imaging can be applied for investigation of structural changes in connective tissue due to various diseases and is beneficial in observing ECM and cell nuclei in cancer diagnostics and prognostics.

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Mykolas Maciulis, Viktoras Mazeika, Vilte G. Samsone, Lukas Kontenis, Edvardas Zurauskas, Vitalijus Karabanovas, and Virginijus Barzda "Polarimetric third-harmonic generation microscopy for histopathology", Proc. SPIE PC12846, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XXII, PC128460D (13 March 2024); https://doi.org/10.1117/12.3001641

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