Precise nanosizing with high dynamic range holography

TL;DR

This paper introduces off-axis k-space holography, a novel imaging technique that significantly enhances dynamic range for precise nanoscale sizing of particles and extracellular vesicles in optical sensing.

Contribution

The authors develop and validate a new holography method that overcomes dynamic range limitations, enabling accurate size measurement of nanometric particles in complex biological samples.

Findings

Achieved up to 110 dB dynamic range in imaging.

Successfully sized metallic and dielectric particles across a large field-of-view.

Provided quantitative size-distributions of extracellular vesicles.

Abstract

Optical sensing is one of the key-enablers of modern diagnostics. Especially label-free imaging modalities hold great promise as they eliminate labeling procedures prior to analysis. However, scattering signals of nanometric particles scale with their volume-square. This unfavorable scaling makes it extremely difficult to quantitatively characterize intrinsically heterogeneous clinical samples, such as extracellular vesicles, as their signal variation easily exceeds the dynamic range of currently available cameras. Here, we introduce off-axis k-space holography that circumvents this limitation. By imaging the back-focal-plane of our microscope we project the scattering signal of all particles onto all camera pixels thus dramatically boosting the achievable dynamic range to up-to 110 dB. We validate our platform by detecting, and quantitatively sizing, metallic and dielectric particles over a 200x200 $\mu$m field-of-view and demonstrate that independently performed signal calibrations allow correctly sizing particles made from different materials. Finally, we present quantitative size-distributions of extracellular vesicle samples.