title: Droplet microfluidic technology for single-cell high-throughput screening.
abstract: We present a droplet-based microfluidic technology that enables high-throughput screening of single mammalian cells. This integrated platform allows for the encapsulation of single cells and reagents in independent aqueous microdroplets (1 pL to 10 nL volumes) dispersed in an immiscible carrier oil and enables the digital manipulation of these reactors at a very high-throughput. Here, we validate a full droplet screening workflow by conducting a droplet-based cytotoxicity screen. To perform this screen, we first developed a droplet viability assay that permits the quantitative scoring of cell viability and growth within intact droplets. Next, we demonstrated the high viability of encapsulated human monocytic U937 cells over a period of 4 days. Finally, we developed an optically-coded droplet library enabling the identification of the droplets composition during the assay read-out. Using the integrated droplet technology, we screened a drug library for its cytotoxic effect against U937 cells. Taken together our droplet microfluidic platform is modular, robust, uses no moving parts, and has a wide range of potential applications including high-throughput single-cell analyses, combinatorial screening, and facilitating small sample analyses.
authors.affiliation: RainDance Technologies, Lexington, MA 02421, USA. brouzese@raindancetech.com
authors.forename: Eric
authors.initials: E
authors.lastname: Brouzes
authors.forename: Martina
authors.initials: M
authors.lastname: Medkova
authors.forename: Neal
authors.initials: N
authors.lastname: Savenelli
authors.forename: Dave
authors.initials: D
authors.lastname: Marran
authors.forename: Mariusz
authors.initials: M
authors.lastname: Twardowski
authors.forename: J Brian
authors.initials: JB
authors.lastname: Hutchison
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Rothberg
authors.forename: Darren R
authors.initials: DR
authors.lastname: Link
authors.forename: Norbert
authors.initials: N
authors.lastname: Perrimon
authors.forename: Michael L
authors.initials: ML
authors.lastname: Samuels
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004655:Emulsions; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000379:methods; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D016685:Mitomycin / Q000737:chemistry / Q000494:pharmacology; D015203:Reproducibility of Results; D013997:Time Factors; D020298:U937 Cells
chemical_list: D004655:Emulsions; D005456:Fluorescent Dyes; D016685:Mitomycin