EPD-iSCAT: Electrophoretic Mass Photometry

25 October 2023, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Interferometric scattering microscopy (iSCAT) has rapidly developed as a quantitative tool for the label-free detection of single macromolecules and nanoparticles. In practice, this measurement records the interferometric scattering signal of individual nanoparticles in solution as they land and stick on a coverslip, exhibiting an intensity that varies linearly with particle volume, and an adsorption rate that reflects the solution-phase transport kinetics of the system. Together, such measurements provide a multidimensional gauge of particle size and concentration in solution over time. However, the landing kinetics of particles in solution also manifest a measurement frequency limitation imposed by the slow long-range mobility of particle diffusion to the measurement interface. Here we introduce a new technique that offers a novel means to overcome the inherent diffusion-controlled sampling limitation of spontaneous mass photometry. We term this methodology, electrophoretic deposition interferometric scattering microscopy (EPD-iSCAT). This approach uses a coverslip supporting a conductive thin film of indium tin oxide (ITO). Charging this ITO film to a potential of around 1 V electrophoretically draws charged nanoparticles from solution and binds them in the focal plane of the microscope. Regulating this potential offers a direct means to control particle deposition. Thus, we find for a 0.1 nM solution of 50 nm polystyrene nanoparticles that the application of +1 V to an EPD-iSCAT coverslip assembly drives a electrophoetic deposition rate constant of 1.7 s−1 μm−2 nM−1. Removal of the potential causes deposition to cease. This user control of EPD-iSCAT affords a means to apply single-molecule mass photometery to monitor long-term changes in solution owing to slow kinetic processes. In contrast with conventional coverslips chemically derivatized with charged thin films, EPD-iSCAT maintains a deposition rate that varies linearly with bulk concentration.

Keywords

iSCAT
Mass Photometry
Indium Tin Oxide
Nanoplastics

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Additional details, specifications and discussion regarding device construction, image classification, metrics and limitations.
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