Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing

Jack Harrington; Luis Blay Esteban; Jonathan Butement; Andres F. Vallejo; Simon I. R. Lane; Bhavwanti Sheth; Maaike S. A. Jongen; Rachel Parker; Patrick S. Stumpf; Rosanna C. G. Smith; Ben D. MacArthur; Matthew J. J. Rose-Zerilli; Marta E. Polak; Tim Underwood; Jonathan West

doi:10.1039/D1LC00292A

Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing†

Jack Harrington,

^a Luis Blay Esteban,

^b Jonathan Butement,^a Andres F. Vallejo,

^c Simon I. R. Lane,^bd Bhavwanti Sheth,^e Maaike S. A. Jongen,

^a Rachel Parker,^a Patrick S. Stumpf,^f Rosanna C. G. Smith,^a Ben D. MacArthur,^dg Matthew J. J. Rose-Zerilli,^ad Marta E. Polak,

^cd Tim Underwood^ad and Jonathan West

*^ad

Author affiliations

* Corresponding authors

^a Cancer Sciences, Faculty of Medicine, University of Southampton, UK
E-mail: J.J.West@soton.ac.uk

^b Faculty of Engineering and Physical Sciences, University of Southampton, UK

^c Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, Faculty of Medicine, University of Southampton, UK

^d Institute for Life Sciences, University of Southampton, UK

^e School for Biological Sciences, Faculty of Environmental and Life Sciences, University of Southampton, UK

^f Centre for Human Development, Stem Cells and Regeneration, Faculty of Medicine, University of Southampton, UK

^g Mathematical Sciences, University of Southampton, UK

Abstract

The future of single cell diversity screens involves ever-larger sample sizes, dictating the need for higher throughput methods with low analytical noise to accurately describe the nature of the cellular system. Current approaches are limited by the Poisson statistic, requiring dilute cell suspensions and associated losses in throughput. In this contribution, we apply Dean entrainment to both cell and bead inputs, defining different volume packets to effect efficient co-encapsulation. Volume ratio scaling was explored to identify optimal conditions. This enabled the co-encapsulation of single cells with reporter beads at rates of ∼1 million cells per hour, while increasing assay signal-to-noise with cell multiplet rates of ∼2.5% and capturing ∼70% of cells. The method, called Pirouette coupling, extends our capacity to investigate biological systems.

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Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing†

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Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing

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