Multiscale Modeling and Analysis for High-fidelity Interferometric Scattering Microscopy

Interferometric scattering microscopy (iSCAT), as an ultrasensitive fluorescence-free imaging modality, has recently gain enormous attention and been rapidly developing from demonstration of principle to quantitative sensing. Here we report on a theoretical and experimental study for iSCAT with samples having structural dimensions that differ by 4-5 orders of magnitude. In particular, we demonstrate and intuitively explain the profound effects of sub-nanometer surface roughness of a glass coverslip and of a mica surface on the absolute signal and the shape of the point spread function of a gold nanoparticle. These quantities significantly affect the accuracies for determining the target size and position in all three dimensions. Moreover, we investigate a sample system mimicking a gold nanoparticle in a simplified cell environment and show position-dependent and even asymmetric point spread function of the nanoparticle. The multiscale study will facilitate the development of high fidelity iSCAT in real applications.

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