Dual-angle interferometric scattering microscopy for optical multiparametric particle characterization

Traditional single-nanoparticle sizing using optical microscopy techniques assesses size via the diffusion constant, which requires suspended particles in a medium of known viscosity. However, these assumptions are typically not fulfilled in complex natural sample environments. Here, we introduce dual-angle interferometric scattering microscopy (DAISY), enabling optical quantification of both size and polarizability of individual nanoparticles without requiring a priori information regarding the surrounding media or super-resolution imaging. DAISY achieves this by combining the information contained in concurrently measured forward and backward scattering images through twilight off-axis holography and interferometric scattering (iSCAT). Going beyond particle size and polarizability, single-particle morphology can be deduced from the fact that hydrodynamic radius relates to the outer particle radius while the scattering-based size estimate depends on the internal mass distribution of the particles. We demonstrate this by optically differentiating biomolecular fractal aggregates from spherical particles in fetal bovine serum at the single particle level.