{"doi":"10.1039/d0lc00261e","title":"Double emulsion flow cytometry with high-throughput single droplet isolation and nucleic acid recovery","abstract":"Droplet microfluidics has made large impacts in diverse areas such as enzyme evolution, chemical product screening, polymer engineering, and single-cell analysis. However, while droplet reactions have become increasingly sophisticated, phenotyping droplets by a fluorescent signal and sorting them to isolate individual variants-of-interest at high-throughput remains challenging. Here, we present sdDE-FACS (s[combining low line]ingle d[combining low line]roplet D[combining low line]ouble E[combining low line]mulsion-FACS), a new method that uses a standard flow cytometer to phenotype, select, and isolate individual double emulsion droplets of interest. Using a 130 μm nozzle at high sort frequency (12-14 kHz), we demonstrate detection of droplet fluorescence signals with a dynamic range spanning 5 orders of magnitude and robust post-sort recovery of intact double emulsion (DE) droplets using 2 commercially-available FACS instruments. We report the first demonstration of single double emulsion droplet isolation with post-sort recovery efficiencies >70%, equivalent to the capabilities of single-cell FACS. Finally, we establish complete downstream recovery of nucleic acids from single, sorted double emulsion droplets via qPCR with little to no cross-contamination. sdDE-FACS marries the full power of droplet microfluidics with flow cytometry to enable a variety of new droplet assays, including rare variant isolation and multiparameter single-cell analysis.","journal":"Lab on a Chip","year":2020,"id":54253,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":106,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9615,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":280070,"name":"Catherine Carswell-Crumpton","orcid":null,"position":1,"is_corresponding":false},{"id":278291,"name":"Sandy Klemm","orcid":"0000-0001-7556-6795","position":2,"is_corresponding":false},{"id":278292,"name":"Bianca Cruz","orcid":"0000-0003-3914-4936","position":3,"is_corresponding":false},{"id":278293,"name":"Gaeun Kim","orcid":"0009-0008-1502-1847","position":4,"is_corresponding":false},{"id":278294,"name":"Suzanne G. K. Calhoun","orcid":"0000-0003-3368-6881","position":5,"is_corresponding":false},{"id":278295,"name":"Lisa Nichols","orcid":"0000-0003-3635-0435","position":6,"is_corresponding":false},{"id":278296,"name":"Polly M. Fordyce","orcid":"0000-0002-9505-0638","position":7,"is_corresponding":false},{"id":280069,"name":"Kara K. Brower","orcid":null,"position":0,"is_corresponding":true}],"reference_count":63,"raw_metadata":null,"created_at":"2026-07-18T21:03:43.037640Z","pmid":"32417874","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}