Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing
Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing
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.
3378-3386
Harrington, Jack
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Esteban, Luis Blay
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Butement, Jonathan
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Vallejo, Andres F
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Lane, Simon I R
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Sheth, Bhavwanti
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Jongen, Maaike S A
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Parker, Rachel
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Stumpf, Patrick S
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Smith, Rosanna C G
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MacArthur, Ben D
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Rose-Zerilli, Matthew J J
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Polak, Marta E
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Underwood, Tim
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West, Jonathan
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30 June 2021
Harrington, Jack
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Esteban, Luis Blay
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Butement, Jonathan
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Vallejo, Andres F
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Lane, Simon I R
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Sheth, Bhavwanti
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Jongen, Maaike S A
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Parker, Rachel
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Stumpf, Patrick S
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Smith, Rosanna C G
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MacArthur, Ben D
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Rose-Zerilli, Matthew J J
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Polak, Marta E
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Underwood, Tim
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West, Jonathan
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Harrington, Jack, Esteban, Luis Blay, Butement, Jonathan, Vallejo, Andres F, Lane, Simon I R, Sheth, Bhavwanti, Jongen, Maaike S A, Parker, Rachel, Stumpf, Patrick S, Smith, Rosanna C G, MacArthur, Ben D, Rose-Zerilli, Matthew J J, Polak, Marta E, Underwood, Tim and West, Jonathan
(2021)
Dual dean entrainment with volume ratio modulation for efficient droplet co-encapsulation: extreme single-cell indexing.
Lab on a Chip, 21 (17), .
(doi:10.1039/d1lc00292a).
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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e-pub ahead of print date: 30 June 2021
Published date: 30 June 2021
Additional Information:
Funding Information:
We are indebted to Deborah Mackay for her support. We also thank Gessa Sugiyarto for preparing cells. The project was made possible by financial contributions from the Medical Research Council (JB and MSAJ: MC_PC_15078), a CRUK & Royal College of Surgeons of England Advanced Clinician Scientist Fellowship (JH and TU), an EPSRC CASE studentship (LBE, 1658462), GSK (AFV, ARCP006668), a Sir Henry Dale Wellcome Trust Fellowship (MEP; 109377/Z/15/Z) and a John Goldman Fellowship (MJJRZ; 2016/JGF/0003).
Publisher Copyright:
© 2021 The Royal Society of Chemistry.
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
Identifiers
Local EPrints ID: 450486
URI: http://eprints.soton.ac.uk/id/eprint/450486
ISSN: 1473-0197
PURE UUID: 9a20e15f-f2ee-4ac7-8257-de2233ff28a7
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Date deposited: 29 Jul 2021 16:32
Last modified: 30 Oct 2023 02:59
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Contributors
Author:
Jack Harrington
Author:
Jonathan Butement
Author:
Andres F Vallejo
Author:
Simon I R Lane
Author:
Maaike S A Jongen
Author:
Rachel Parker
Author:
Patrick S Stumpf
Author:
Rosanna C G Smith
Author:
Matthew J J Rose-Zerilli
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