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Traditional single-nanoparticle sizing using optical microscopy techniquesassesses size via the diffusion constant, which requires suspended particles ina medium of known viscosity. However, these assumptions are typically notfulfilled in complex natural sample environments. Here, we introduce dual-angleinterferometric scattering microscopy (DAISY), enabling optical quantificationof both size and polarizability of individual nanoparticles without requiring apriori information regarding the surrounding media or super-resolution imaging.DAISY achieves this by combining the information contained in concurrentlymeasured forward and backward scattering images through twilight off-axisholography and interferometric scattering (iSCAT). Going beyond particle sizeand polarizability, single-particle morphology can be deduced from the factthat hydrodynamic radius relates to the outer particle radius while thescattering-based size estimate depends on the internal mass distribution of theparticles. We demonstrate this by optically differentiating biomolecularfractal aggregates from spherical particles in fetal bovine serum at the singleparticle level.

Dual-angle interferometric scattering microscopy for optical multiparametric particle characterization