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Ultrasonically-Guided Flow Focusing Generates Precise Emulsion Droplets for High-Throughput Single Cell Analyses Short Title: Precision Droplet Generation for Single Cell Analysis

Author:
Lagerman, Colton E., López Acevedo, Sheila N., Fahad, Ahmed S., Hailemariam, Amen T., Madan, Bharat, DeKosky, Brandon J.
Source:
Journal of bioscience and bioengineering 2019
ISSN:
1389-1723
Subject:
B-lymphocytes, antibodies, bioengineering, biotechnology, droplet size, droplets, emulsions, equipment, flow cytometry, genetic analysis, instrumentation, microfluidic technology, ultrasonics, vibration
Abstract:
Emulsion-based techniques have dramatically advanced our understanding of single-cell biology and complex single-cell features over the past two decades. Most approaches for precise single cell isolation rely on microfluidics, which has proven highly effective but requires substantial investment in equipment and expertise that can be difficult to access for researchers that specialize in other areas of bioengineering and molecular biotechnology. Inspired by the robust droplet generation technologies in modern flow cytometry instrumentation, here we established a new platform for high-throughput isolation of single cells within droplets of tunable sizes by combining flow focusing with ultrasonic vibration for rapid and effective droplet formation. Application of ultrasonic pressure waves to the flowing jet provided enhanced control of emulsion droplet size, permitting capture of 25,000 to 50,000 single cells per minute. As an example application, we applied this new droplet generation platform to sequence the antibody variable region heavy and light chain pairings (VH:VL) from large repertoires of single B cells. We demonstrated the recovery of >40,000 paired CDRH3:CDRL3 antibody clusters from a single individual, validating that these droplet systems can enable the genetic analysis of very large single-cell populations. These accessible new technologies will allow rapid, large-scale, and precise single-cell analyses for a broad range of bioengineering and molecular biotechnology applications.
Agid:
6342150