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Optical diffraction tomography for assessing single cell models in angular light scattering

preprint
posted on 2023-11-16, 08:21 authored by Andrew Berger, Kaitlin Dunn, Alex Matlock, George Funkenbusch, Zahid Yaqoob, Peter So
Angularly resolved light scattering (ALS) has become a useful tool for assessing size and refractive index of biological scatterers at cellular and organelle length scales. Sizing organelle populations with ALS relies on Mie scattering theory models, which require significant assumptions about the object, including spherical scatterers and a homogeneous medium. These assumptions may incur greater error at the single cell level, where there are fewer scatterers to be averaged over. We investigate the validity of these assumptions using 3D refractive index (RI) tomograms measured via optical diffraction tomography (ODT). We compute the angular scattering on digitally manipulated tomograms with increasingly strong model assumptions including RI-matched immersion media, homogeneous cytosol, and spherical organelles. We also compare the tomogram-computed angular scattering to experimental measurements of angular scattering from the same cells to ensure that the ODT-based approach accurately models angular scattering. We show that enforced RI-matching with the immersion medium and a homogeneous cytosol significantly affect the angular scattering intensity shape, suggesting that these assumptions can reduce the accuracy of size distribution estimates.

History

Funder Name

National Institutes of Health (P41EB015871,R01DA045549,R21GM140613-02,R01HL158102); Hamamatsu Corporation; The MIT Laser Biomedical Research Center

Preprint ID

110812

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