Scalable Fabrication and Use of 3D Structured Microparticles Spatially Functionalized with Biomolecules

Sohyung Lee, Joseph de Rutte, Robert Dimatteo, Doyeon Koo, and Dino Di Carlo

ACS Nano, Nov 2021

DOI: 10.1021/acsnano.1c05857   |   PubMed: 34846855   |   bioRxiv


Abstract

Microparticles with defined shapes and spatial chemical modification can interface with cells and tissues at the cellular scale. However, conventional methods to fabricate shaped microparticles have trade-offs between the throughput of manufacture and the precision of particle shape and chemical functionalization. Here, we achieved scalable production of hydrogel microparticles at rates of greater than 40 million/hour with localized surface chemistry using a parallelized step emulsification device and temperature-induced phase-separation. The approach harnesses a polymerizable polyethylene glycol (PEG) and gelatin aqueous two-phase system (ATPS) which conditionally phase separates within microfluidically generated droplets. Following droplet formation, phase separation is induced and phase separated droplets are subsequently cross-linked to form uniform crescent and hollow shell particles with gelatin functionalization on the boundary of the cavity. The gelatin localization enabled deterministic cell loading in subnanoliter-sized crescent-shaped particles, which we refer to as nanovials, with cavity dimensions tuned to the size of cells. Loading on nanovials also imparted improved cell viability during analysis and sorting using standard fluorescence activated cell sorters, presumably by protecting cells from shear stress. This localization effect was further exploited to selectively functionalize capture antibodies to nanovial cavities enabling single-cell secretion assays with reduced cross-talk in a simplified format.

 

Topics

Technology, Workflow Improvement

Cell Types

CHO Cells

Flow Cytometers

SONY SH800

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