Machine learning enables precise holographic characterization of colloidal materials in real time.

Holographic particle characterization uses in-line holographic video microscopy to track and characterize individual colloidal particles dispersed in their native fluid media. Applications range from fundamental research in statistical physics to product development in biopharmaceuticals and medical diagnostic testing. The information encoded in a hologram can be extracted by fitting to a generative model based on the Lorenz-Mie theory of light scattering. Treating hologram analysis as a high-dimensional inverse problem has been exceptionally successful, with conventional optimization algorithms yielding nanometer precision for a typical particle's position and part-per-thousand precision for its size and index of refraction. Machine learning previously has been used to automate holographic particle characterization by detecting features of interest in multi-particle holograms and estimating the particles' positions and properties for subsequent refinement. This study presents an updated end-to-end neural-network solution called CATCH (Characterizing and Tracking Colloids Holographically) whose predictions are fast, precise, and accurate enough for many real-world high-throughput applications and can reliably bootstrap conventional optimization algorithms for the most demanding applications. The ability of CATCH to learn a representation of Lorenz-Mie theory that fits within a diminutive 200 kB hints at the possibility of developing a greatly simplified formulation of light scattering by small objects.

Multi-angle holographic characterization of individual fractal aggregates.

Holographic particle characterization uses quantitative analysis of holographic microscopy data to precisely and rapidly measure the diameter and refractive index of individual colloidal spheres in their native media. When this technique is applied to inhomogeneous or aspherical particles, the measured diameter and refractive index represent properties of an effective sphere enclosing each particle. Effective-sphere analysis has been applied successfully to populations of fractal aggregates, yielding an overall fractal dimension for the population as a whole. Here, we demonstrate that holographic characterization also can measure the fractal dimensions of an individual fractal cluster by probing how its effective diameter and refractive index change as it undergoes rotational diffusion. This procedure probes the structure of a cluster from multiple angles and thus constitutes a form of tomography. Here we demonstrate and validate this effective-sphere interpretation of aspherical particles' holograms through experimental studies on aggregates of silica nanoparticles grown under a range of conditions.

Holographic characterization and tracking of colloidal dimers in the effective-sphere approximation.

An in-line hologram of a colloidal sphere can be analyzed with the Lorenz-Mie theory of light scattering to measure the sphere's three-dimensional position with nanometer-scale precision while also measuring its diameter and refractive index with part-per-thousand precision. Applying the same technique to aspherical or inhomogeneous particles yields measurements of the position, diameter and refractive index of an effective sphere that represents an average over the particle's geometry and composition. This effective-sphere interpretation has been applied successfully to porous, dimpled and coated spheres, as well as to fractal clusters of nanoparticles, all of whose inhomogeneities appear on length scales smaller than the wavelength of light. Here, we combine numerical and experimental studies to investigate effective-sphere characterization of symmetric dimers of micrometer-scale spheres, a class of aspherical objects that appear commonly in real-world dispersions. Our studies demonstrate that the effective-sphere interpretation usefully distinguishes small colloidal clusters in holographic characterization studies of monodisperse colloidal spheres. The effective-sphere estimate for a dimer's axial position closely follows the ground truth for its center of mass. Trends in the effective-sphere diameter and refractive index, furthermore, can be used to measure a dimer's three-dimensional orientation. When applied to colloidal dimers transported in a Poiseuille flow, the estimated orientation distribution is consistent with expectations for Brownian particles undergoing Jeffery orbits.

Anomalous tumbling of colloidal ellipsoids in Poiseuille flows.

Shear flows cause aspherical colloidal particles to tumble so that their orientations trace out complex trajectories known as Jeffery orbits. The Jeffery orbit of a prolate ellipsoid is predicted to align the particle's principal axis preferentially in the plane transverse to the axis of shear. Holographic microscopy measurements reveal instead that colloidal ellipsoids' trajectories in Poiseuille flows strongly favor an orientation inclined by roughly π/8 relative to this plane. This anomalous observation is consistent with at least two previous reports of colloidal rods and dimers of colloidal spheres in Poiseuille flow and therefore appears to be a generic, yet unexplained feature of colloidal transport at low Reynolds numbers.

Holographic analysis of colloidal spheres sedimenting in horizontal slit pores.

The mobility of a colloidal particle in a slit pore is modified by the particle's hydrodynamic coupling to the bounding surfaces and therefore depends on the particle's position within the pore and its direction of motion. We report holographic particle tracking measurements of colloidal particles' diffusion and sedimentation between parallel horizontal walls that yield the mobility for motions perpendicular to the walls, including its dependence on height within the channel. These measurements complement previous studies that probed colloidal mobility parallel to confining surfaces. When interpreted with effective-medium theory, holographic characterization measurements yield estimates for the sedimenting spheres' densities that can be compared with kinematic values to draw insights into the spheres' compositions. This comparison suggests, for example, that the silica spheres used in this study are slightly porous, but that their pores are too small for water to penetrate.

Interpreting holographic molecular binding assays with effective medium theory.

Holographic molecular binding assays use holographic video microscopy to directly detect molecules binding to the surfaces of micrometer-scale colloidal beads by monitoring associated changes in the beads' light-scattering properties. Holograms of individual spheres are analyzed by fitting to a generative model based on the Lorenz-Mie theory of light scattering. Each fit yields an estimate of a probe bead's diameter and refractive index with sufficient precision to watch a population of beads grow as molecules bind. Rather than modeling the molecular-scale coating, however, these fits use effective medium theory, treating the coated sphere as if it were homogeneous. This effective-sphere analysis is rapid and numerically robust and so is useful for practical implementations of label-free immunoassays. Here, we assess how measured effective-sphere properties reflect the actual properties of molecular-scale coatings by modeling coated spheres with the discrete-dipole approximation and analyzing their holograms with the effective-sphere model.

CATCH: Characterizing and Tracking Colloids Holographically Using Deep Neural Networks.

In-line holographic microscopy provides an unparalleled wealth of information about the properties of colloidal dispersions. Analyzing one colloidal particle's hologram with the Lorenz-Mie theory of light scattering yields the particle's three-dimensional position with nanometer precision while simultaneously reporting its size and refractive index with part-per-thousand resolution. Analyzing a few thousand holograms in this way provides a comprehensive picture of the particles that make up a dispersion, even for complex multicomponent systems. All of this valuable information comes at the cost of three computationally expensive steps: (1) identifying and localizing features of interest within recorded holograms, (2) estimating each particle's properties based on characteristics of the associated features, and finally (3) optimizing those estimates through pixel-by-pixel fits to a generative model. Here, we demonstrate an end-to-end implementation that is based entirely on machine-learning techniques. Characterizing and Tracking Colloids Holographically (CATCH) with deep convolutional neural networks is fast enough for real-time applications and otherwise outperforms conventional analytical algorithms, particularly for heterogeneous and crowded samples. We demonstrate this system's capabilities with experiments on free-flowing and holographically trapped colloidal spheres.