期刊
ACS NANO
卷 16, 期 1, 页码 38-49出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.1c05857
关键词
microfluidics; microparticle; ATPS; single-cell analysis; flow cytometry
类别
资金
- NIH Shared Instrumentation Grant [S10OD025017]
- NSF [CHE-0722519]
- UCLA W. M. Keck Foundation COVID 19 Research Award Program
- National Institutes of Health [P30 CA016042, 5P30 AI028697]
- JCCC
- David Geffen School of Medicine at UCLA
- UCLA Chancellor's Office
- UCLA Vice Chancellor's Office of Research
- UCLA AIDS Institute
Researchers have developed a scalable method for manufacturing hydrogel microparticles with defined shapes and chemical functionalization. The process involves using a two-phase system and microfluidic technology, and allows for localized surface chemistry on the microparticles. These microparticles can be used for cell loading and single-cell secretion analysis.
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.
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