Double Emulsion Picoreactors for High-Throughput Single-Cell Encapsulation and Phenotyping via FACS

• Published in: Analytical chemistry (Washington) 2020-10, Vol.92 (19), p.13262-13270
• Main Authors: Brower, Kara K, Khariton, Margarita, Suzuki, Peter H, Still, Chris, Kim, Gaeun, Calhoun, Suzanne G. K, Qi, Lei S, Wang, Bo, Fordyce, Polly M
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Animals; Biotechnology; Cell Encapsulation; Cell Line; Cell lines; Cell size; Chemistry; Diameters; Double emulsions; Droplets; Emulsions; Encapsulation; Flow Cytometry; Fluorescence; Genetic analysis; Genomes; Genotypes; High-throughput screening; High-Throughput Screening Assays; Mice; Microfluidic Analytical Techniques; Microfluidics; Morphology; Nucleic acids; Particle Size; Phenotype; Phenotypes; Phenotyping; Robustness; Screening; Single-Cell Analysis; Surface Properties; Workflow
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.0c02499

In the past five years, droplet microfluidic techniques have unlocked new opportunities for the high-throughput genome-wide analysis of single cells, transforming our understanding of cellular diversity and function. However, the field lacks an accessible method to screen and sort droplets based on cellular phenotype upstream of genetic analysis, particularly for large and complex cells. To meet this need, we developed Dropception, a robust, easy-to-use workflow for precise single-cell encapsulation into picoliter-scale double emulsion droplets compatible with high-throughput screening via fluorescence-activated cell sorting (FACS). We demonstrate the capabilities of this method by encapsulating five standardized mammalian cell lines of varying sizes and morphologies as well as a heterogeneous cell mixture of a whole dissociated flatworm (5–25 μm in diameter) within highly monodisperse double emulsions (35 μm in diameter). We optimize for preferential encapsulation of single cells with extremely low multiple-cell loading events (