Fluorescence-activated droplet sorting (FADS): efficient microfluidic cell sorting based on enzymatic activity

Jean-Christophe Baret; Oliver J. Miller; Valerie Taly; Michaël Ryckelynck; Abdeslam El-Harrak; Lucas Frenz; Christian Rick; Michael L. Samuels; J. Brian Hutchison; Jeremy J. Agresti; Darren R. Link; David A. Weitz; Andrew D. Griffiths

doi:10.1039/B902504A

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Lab on a Chip

Issue 13, 2009

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Fluorescence-activated droplet sorting (FADS): efficient microfluidic cell sorting based on enzymatic activity

Jean-Christophe Baret , Oliver J. Miller , Valerie Taly , Michaël Ryckelynck , Abdeslam El-Harrak , Lucas Frenz , Christian Rick , Michael L. Samuels , J. Brian Hutchison , Jeremy J. Agresti , Darren R. Link , David A. Weitz and Andrew D. Griffiths

Lab Chip, 2009, 9, 1850-1858

DOI: 10.1039/B902504A
Received 05 Feb 2009, Accepted 14 Apr 2009
First published on the web 23 Apr 2009

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We describe a highly efficient microfluidic fluorescence-activated droplet sorter (FADS) combining many of the advantages of microtitre-plate screening and traditional fluorescence-activated cell sorting (FACS). Single cells are compartmentalized in emulsion droplets, which can be sorted using dielectrophoresis in a fluorescence-activated manner (as in FACS) at rates up to 2000 droplets s⁻¹. To validate the system, mixtures of E. coli cells, expressing either the reporter enzyme β-galactosidase or an inactive variant, were compartmentalized with a fluorogenic substrate and sorted at rates of [similar] 300 droplets s⁻¹. The false positive error rate of the sorter at this throughput was <1 in 10⁴ droplets. Analysis of the sorted cells revealed that the primary limit to enrichment was the co-encapsulation of E. coli cells, not sorting errors: a theoretical model based on the Poisson distribution accurately predicted the observed enrichment values using the starting cell density (cells per droplet) and the ratio of active to inactive cells. When the cells were encapsulated at low density ( [similar] 1 cell for every 50 droplets), sorting was very efficient and all of the recovered cells were the active strain. In addition, single active droplets were sorted and cells were successfully recovered.

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