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A programmable droplet-based microfluidic device for multiparameter single-cell analysis

University of British Columbia

The ability of microfluidic systems to perform biological analysis with greater sensitivity, lower cost, and higher throughput relative to conventional methods has now been widely demonstrated. Despite this transformative potential, application innovation and user adoption in biological research have lagged due to limited access to specialized fabrication facilities and expertise. In analogy to how the development of programmable integrated circuits has resulted in the ubiquity and utility of this technology among a broad community of developers and non-expert users, the advancement of programmable microfluidic devices stands to dramatically enhance the pervasiveness and impact of microfluidic systems. This thesis describes the development and application of a microfluidic device that combines the reconfigurable flow-routing capabilities of integrated microvalve technology with the sample compartmentalization inherent to mass transport in droplets to achieve programmable fluidhandling functionality. The device allows for the execution of user-defined multistep reaction protocols in an array of individually addressable nanolitre-volume storage chambers by consecutively merging programmable sequences of picolitre-volume droplets containing reagents or phenotypically sorted single cells. This functionality is enabled by “flow-controlled wetting,” a novel droplet docking and merging mechanism that exploits the physics of droplet flow through a channel to control the precise location of droplet wetting. The device also allows for automated cross-contamination-free recovery of reaction products from individual chambers for downstream analysis. The combined features of programmability, addressability, and selective recovery provide a general hardware platform that can be reprogrammed for multiple applications. This versatility is demonstrated by implementing multiple analyses on phenotypically sorted single cells including monoclonal culture, genomic PCR, whole genome amplification and whole transcriptome amplification. These capabilities have been applied to a diverse range of biological samples for applications ranging from the identification of microbial community members in environmental samples to the determination of mutation frequencies in human cancer at the single-cell level.

Text

2014-01-31

Attribution-NonCommercial-NoDerivatives 4.0 International

http://hdl.handle.net/2429/43855

Electrical and Computer Engineering

University of British Columbia

UBCV

http://creativecommons.org/licenses/by-nc-nd/4.0/