Wide dynamic range imaging with bidirectional quantitative scattering microscopy

Kohki Horie; Keiichiro Toda; Takuma Nakamura; Takuro Ideguchi

doi:10.1117/12.3000957

12 March 2024 Wide dynamic range imaging with bidirectional quantitative scattering microscopy

Kohki Horie, Keiichiro Toda, Takuma Nakamura, Takuro Ideguchi

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Proceedings Volume 12852, Quantitative Phase Imaging X; 128520G (2024) https://doi.org/10.1117/12.3000957
Event: SPIE BiOS, 2024, San Francisco, California, United States

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Abstract

Quantitative phase microscopy (QPM), which quantifies the complex amplitude of forward-scattered (FS) light induced by specimens, enables the visualization of the macroscopic refractive index distribution in transparent samples and has been used for various applications such as dry mass analysis and cell cycle estimation. Conversely, interferometric scattering microscopy (iSCAT), which captures backward-scattered (BS) light, offers higher sensitivity compared to QPM and can effectively detect nanoparticles such as individual proteins and viruses, with the drawback of being unable to provide quantitative information on macrostructure. These microscopy techniques provide complementary information and have undergone independent advancements. In this study, we present bidirectional quantitative scattering microscopy (bidirectional QSM) for wide dynamic range imaging by simultaneously performing FS and BS measurements with a single image sensor using a digital holographic technique. This approach enables quantitative measurement of macrostructures as well as high sensitivity to nanoparticles. Moreover, digital holography facilitates the acquisition of complex amplitude FS and BS images, which allows simultaneous computational z-axis focusing, enabling subsequent quantitative correlation analysis between them. To provide a practical demonstration, time-lapse imaging of a cell is performed to characterize temporal changes in the state of cellular vital activity.

(2024) Published by SPIE. Downloading of the abstract is permitted for personal use only.

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Kohki Horie, Keiichiro Toda, Takuma Nakamura, and Takuro Ideguchi "Wide dynamic range imaging with bidirectional quantitative scattering microscopy", Proc. SPIE 12852, Quantitative Phase Imaging X, 128520G (12 March 2024); https://doi.org/10.1117/12.3000957

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