A programmable droplet-based microfluidic device applied to multiparameter analysis of single microbes and microbial communities

Proc Natl Acad Sci U S A. 2012 May 15;109(20):7665-70. doi: 10.1073/pnas.1106752109. Epub 2012 Apr 30.

Abstract

We present a programmable droplet-based microfluidic device that combines the reconfigurable flow-routing capabilities of integrated microvalve technology with the sample compartmentalization and dispersion-free transport that is inherent to droplets. The device allows for the execution of user-defined multistep reaction protocols in 95 individually addressable nanoliter-volume storage chambers by consecutively merging programmable sequences of picoliter-volume droplets containing reagents or cells. This functionality is enabled by "flow-controlled wetting," a 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 into standard microfuge tubes for downstream analysis. The combined features of programmability, addressability, and selective recovery provide a general hardware platform that can be reprogrammed for multiple applications. We demonstrate this versatility by implementing multiple single-cell experiment types with this device: bacterial cell sorting and cultivation, taxonomic gene identification, and high-throughput single-cell whole genome amplification and sequencing using common laboratory strains. Finally, we apply the device to genome analysis of single cells and microbial consortia from diverse environmental samples including a marine enrichment culture, deep-sea sediments, and the human oral cavity. The resulting datasets capture genotypic properties of individual cells and illuminate known and potentially unique partnerships between microbial community members.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Primers / genetics
  • Genotype
  • Geologic Sediments / microbiology
  • Humans
  • Hydrodynamics*
  • Image Processing, Computer-Assisted
  • Metagenome / genetics*
  • Metagenomics / methods
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods*
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Mouth / microbiology
  • Polymerase Chain Reaction
  • RNA, Ribosomal, 16S / genetics
  • Sequence Analysis, DNA
  • Surface-Active Agents
  • Wettability

Substances

  • DNA Primers
  • RNA, Ribosomal, 16S
  • Surface-Active Agents