Microfluidic Platforms for Medical Diagnostics & Synthetic Biology Applications

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Author(s)
Singh, Anup K.
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School of Chemical and Biomolecular Engineering
School established in 1901 as the School of Chemical Engineering; in 2003, renamed School of Chemical and Biomolecular Engineering
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Abstract
Microfluidic assays and devices have attracted a significant attention for performing biochemical reactions and analysis as they provide significant improvements over their macroscale counterparts with respect to speed, throughput, sensitivity and multiplexing. We are involved in developing innovative microfluidic assays and integrated devices for many applications including cell signaling, systems biology, medical diagnostics and synthetic biology. In this talk, I would present application of microfluidic platforms in two areas--medical diagnostics and synthetic biology. The first example describes the development of a centrifugal microfluidic device (SpinDx) capable of analyzing clinical (e.g., whole blood) samples for multiplexed diagnostics. The device is targeted for point-of-care applications and is rapid (<15-30 min sample-to-answer), sensitive (sub-pM detection limit), multiplexed (up to 64 parallel assays), and capable of analyzing minute sample amounts (< 20 μL total sample input). The immunoassay approach is based on binding of an analyte in sample to antibody-laden capture particles followed by sedimentation of particles through a density-media in a microfluidic disk and quantification using a laser-induced fluorescence detector. A key advantage of the technique is its compatibility with a variety of sample matrices with no off-disk sample preparation required. We have demonstrated detection of many analytes including biotoxins, pathogenic bacteria, and immune-response markers in exogeneous whole blood, exogeneous and endogeneous serum, saliva and environmental samples. The second example consists of a droplet microfluidic platform for significantly improving the throughput and reducing the cost of synthetic biology experiments. Synthetic biology experiments require optimization of pathways consisting of many genes and other genetic elements and given the large number of alternatives available for each element, optimization of a pathway can require very large number of experiments. Currently, these experiments are done manually using fairly large amount of costly reagents per experiment making the process very expensive, extremely slow and irreproducible. We have developed a platform that uses droplets as discrete reaction chambers to integrate and automate the processes of DNA assembly, transformation, and cell culture in one device. The hybrid chip combines droplets-in-flow and digital microfluidic (DMF) formats to take advantage of the high throughput nature of droplets-in-flow and the precise control over droplet manipulation offered by the DMF. We show that the platform is capable of accurate DNA assembly, efficient transformation, and cell culture and is compatible with many cloning methods (e.g., Golden Gate and Gibson) and chassis organisms (e.g., bacteria, yeast and fungus).
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Date
2015-11-04
Extent
60:42 minutes
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Moving Image
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Lecture
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