Scalable Device for Automated Microbial Electroporation in a Digital Microfluidic Platform. 2017

Andrew C Madison, and Matthew W Royal, and Frederic Vigneault, and Liji Chen, and Peter B Griffin, and Mark Horowitz, and George M Church, and Richard B Fair
Department of Electrical and Computer Engineering, Duke University , Durham, North Carolina 27708, United States.

Electrowetting-on-dielectric (EWD) digital microfluidic laboratory-on-a-chip platforms demonstrate excellent performance in automating labor-intensive protocols. When coupled with an on-chip electroporation capability, these systems hold promise for streamlining cumbersome processes such as multiplex automated genome engineering (MAGE). We integrated a single Ti:Au electroporation electrode into an otherwise standard parallel-plate EWD geometry to enable high-efficiency transformation of Escherichia coli with reporter plasmid DNA in a 200 nL droplet. Test devices exhibited robust operation with more than 10 transformation experiments performed per device without cross-contamination or failure. Despite intrinsic electric-field nonuniformity present in the EP/EWD device, the peak on-chip transformation efficiency was measured to be 8.6 ± 1.0 × 108 cfu·μg-1 for an average applied electric field strength of 2.25 ± 0.50 kV·mm-1. Cell survival and transformation fractions at this electroporation pulse strength were found to be 1.5 ± 0.3 and 2.3 ± 0.1%, respectively. Our work expands the EWD toolkit to include on-chip microbial electroporation and opens the possibility of scaling advanced genome engineering methods, like MAGE, into the submicroliter regime.

UI MeSH Term Description Entries
D008839 Microelectrodes Electrodes with an extremely small tip, used in a voltage clamp or other apparatus to stimulate or record bioelectric potentials of single cells intracellularly or extracellularly. (Dorland, 28th ed) Electrodes, Miniaturized,Electrode, Miniaturized,Microelectrode,Miniaturized Electrode,Miniaturized Electrodes
D004867 Equipment Design Methods and patterns of fabricating machines and related hardware. Design, Equipment,Device Design,Medical Device Design,Design, Medical Device,Designs, Medical Device,Device Design, Medical,Device Designs, Medical,Medical Device Designs,Design, Device,Designs, Device,Designs, Equipment,Device Designs,Equipment Designs
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D012371 Robotics The application of electronic, computerized control systems to mechanical devices designed to perform human functions. Formerly restricted to industry, but nowadays applied to artificial organs controlled by bionic (bioelectronic) devices, like automated insulin pumps and other prostheses. Companion Robots,Humanoid Robots,Remote Operations (Robotics),Social Robots,Socially Assistive Robots,Telerobotics,Soft Robotics,Assistive Robot, Socially,Companion Robot,Humanoid Robot,Operation, Remote (Robotics),Operations, Remote (Robotics),Remote Operation (Robotics),Robot, Companion,Robot, Humanoid,Robot, Social,Robot, Socially Assistive,Robotic, Soft,Social Robot,Socially Assistive Robot,Soft Robotic
D012815 Signal Processing, Computer-Assisted Computer-assisted processing of electric, ultrasonic, or electronic signals to interpret function and activity. Digital Signal Processing,Signal Interpretation, Computer-Assisted,Signal Processing, Digital,Computer-Assisted Signal Interpretation,Computer-Assisted Signal Interpretations,Computer-Assisted Signal Processing,Interpretation, Computer-Assisted Signal,Interpretations, Computer-Assisted Signal,Signal Interpretation, Computer Assisted,Signal Interpretations, Computer-Assisted,Signal Processing, Computer Assisted
D014162 Transfection The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES. Transfections
D014169 Transformation, Bacterial The heritable modification of the properties of a competent bacterium by naked DNA from another source. The uptake of naked DNA is a naturally occuring phenomenon in some bacteria. It is often used as a GENE TRANSFER TECHNIQUE. Bacterial Transformation
D056656 Lab-On-A-Chip Devices Microdevices that combine microfluidics technology with electrical and/or mechanical functions for analyzing very small fluid volumes. They consist of microchannels etched into substrates made of silicon, glass, or polymer using processes similar to photolithography. The test fluids in the channels can then interact with different elements such as electrodes, photodetectors, chemical sensors, pumps, and valves. Microchip Analytical Devices,Microfluidic Devices,Microfluidic Lab-On-A-Chip,Microfluidic Microchips,Nanochip Analytical Devices,Analytical Device, Microchip,Analytical Device, Nanochip,Analytical Devices, Microchip,Analytical Devices, Nanochip,Device, Lab-On-A-Chip,Device, Microchip Analytical,Device, Microfluidic,Device, Nanochip Analytical,Devices, Lab-On-A-Chip,Devices, Microchip Analytical,Devices, Microfluidic,Devices, Nanochip Analytical,Lab On A Chip Devices,Lab-On-A-Chip Device,Lab-On-A-Chip, Microfluidic,Lab-On-A-Chips, Microfluidic,Microchip Analytical Device,Microchip, Microfluidic,Microchips, Microfluidic,Microfluidic Device,Microfluidic Lab On A Chip,Microfluidic Lab-On-A-Chips,Microfluidic Microchip,Nanochip Analytical Device
D018274 Electroporation A technique in which electric pulses, in kilovolts per centimeter and of microsecond-to-millisecond duration, cause a loss of the semipermeability of CELL MEMBRANES, thus leading to ion leakage, escape of metabolites, and increased uptake by cells of drugs, molecular probes, and DNA. Depending on the dosage, the formation of openings in the cell membranes caused by the electric pulses may or may not be reversible. Electric Field-Mediated Cell Permeabilization,Irreversible Electroporation,Reversible Electroporation,Electropermeabilisation,Electric Field Mediated Cell Permeabilization,Electroporation, Irreversible,Electroporation, Reversible
D019544 Equipment Failure Analysis The evaluation of incidents involving the loss of function of a device. These evaluations are used for a variety of purposes such as to determine the failure rates, the causes of failures, costs of failures, and the reliability and maintainability of devices. Materials Failure Analysis,Prosthesis Failure Analysis,Analysis, Equipment Failure,Analysis, Materials Failure,Analysis, Prosthesis Failure,Analyses, Equipment Failure,Analyses, Materials Failure,Analyses, Prosthesis Failure,Equipment Failure Analyses,Failure Analyses, Equipment,Failure Analyses, Materials,Failure Analyses, Prosthesis,Failure Analysis, Equipment,Failure Analysis, Materials,Failure Analysis, Prosthesis,Materials Failure Analyses,Prosthesis Failure Analyses

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