High Throughput Digital Microfluidics for On-chip Nucleic Acid Detection

Download or read book High Throughput Digital Microfluidics for On-chip Nucleic Acid Detection written by Andrew Carter Hatch and published by . This book was released on 2011 with total page 175 pages. Available in PDF, EPUB and Kindle. Book excerpt: A set of microfluidic design and fabrication methods are put forward as a means to advance digital microfluidics for high throughput on-chip nucleic acid detection. Droplet microfluidic technology enables higher precision processing of Cells and DNA for genotyping, genetic expression, and quantification assays, but is commonly limited in throughput, particularly in terms of real-time analysis. This body of work focuses on methods to enable high-throughput processing of droplets and reactor-arrays with reduced costs and time demands. Four main contributions of this work include methods to rapidly generate, image, and post-process digital reactor arrays, while increasing fluorescence imaging field of view to simultaneously process more reactors at a time, thereby enabling both high-precision and high-throughput analysis. First, a rapid droplet generation and three-dimensional droplet-packing scheme are used to create high-density droplet reactor arrays. This provides higher throughput processing in a smaller area, with as much as three-fold increases in droplet packing density, and 10%-50% increases in area coverage on the imaging sensor. Second, it examines methods to fabricate three-dimensional Micro/Nano well arrays for high throughput Digital PCR processing yielding up to two-fold increases in imaging area-coverage and reactor-array density. Third, a study of three-dimensional reactor array packing is combined with wide-field fluorescence image processing techniques to demonstrate high throughput digital processing of a 12-cm2 area with 21-megapixel resolution for rapid digital PCR quantification. Fourth, it presents a novel technique to selectively sort and isolate droplets of interest based on differences in intrinsic viscous and viscoelastic properties at sorting rates greater than 200 drops/s. The primary findings of these studies and contributions have been published in peer reviewed journals and international conferences. The basis of these findings have been geared to fast and accurate processing of samples containing nucleic acid DNA in a form of digital PCR but can also be applied to other cellular and chemical detection processes. With the fundamental groundwork of developing platforms and techniques to increase throughput of chemical and biological reactions, fields of microfluidics and digital microfluidics can translate from the research workbench to functional and useful platforms.