Droplet Microfluidics for High-Throughput Chemical Analysis and Experimentation

By Robert Kennedy

Chemistry, University of Michigan, Ann Arbor, MI

Published on

Abstract

Manipulating samples as droplets within microfluidic devices has emerged as an interesting approach for chemical analysis and screening. In segmented flow, one embodiment of this technology, nanoliter samples are manipulated in microfluidic channels as plugs separated by an immiscible fluid, such as air or fluorinated oil. These plugs serve as miniature test-tubes in which reactions can be performed at high throughput. Microfluidic tools have been developed to split, dilute, extract, and filter such plugs at rates >10 samples/s. We have developed methods to analyze plug content by electrophoresis and mass spectrometry (MS). A natural application of this technology is for high throughput screening. By coupling droplet manipulation with MS detection, it is possible to greatly reduce reagent consumption and eliminate the need for fluorescent labels or coupled reactions. The technology and application to screens of deacetylase reactions and protein-protein interactions will be presented. A more involved screening allows for monitoring reactions of enzyme variants to identify new biocatalysts. Droplet technology can also be used for chemical monitoring or sensing applications. In this approach samples emerging from a miniaturized sampling device are segmented for later analysis. We have used this method to monitor neurotransmitter dynamics in the brain. The technology and application to studies of neurotransmission in a Huntington’s disease models will be demonstrated.

Bio

Robert Kennedy Robert Kennedy developed an interest in analytical chemistry and chemical separations while earning his BS degree in chemistry at the University of Florida. He became fascinated with the ability of GC to separate subtly different molecules while he was performing undergraduate research in organic chemistry. His analytical classes taught by Prof. John Dorsey further enhanced this interest. He went on to earn a PhD with James Jorgenson at University of North Carolina where is work focused on using open tubular LC to analyze single cells. After a post-doc with Mark Wightman he started his own research program at University of Florida before moving to University of Michigan as the Hobart H. Willard Professor of Chemistry in 2002. His research has combined his lifelong interest in biology with chemical analysis and separations. A theme of his group has been development of miniaturized, high-speed separations for sensing, detection of non-covalent complexes, and screening. His group has developed capillary separation methods for monitoring neurotransmitters in vivo. These methods have been used for studying changes in neurotransmitter concentrations associated with behavior and diseases. His group has also developed microfluidic electrophoresis devices for monitoring insulin secretion from pancreatic β-cells. These methods are coupled with LC-MS metabolomics to understand the biochemical mechanism of insulin secretion and perturbations associated with diabetes. His group is also researching use of rapid electrophoretic and mass spectrometric assays for high-throughput screening. His work has been recognized by several awards including ACS Award in Chromatography, McKnight Award for Technical Innovations in Neuroscience, EAS Separation Science Award, Golay Award for Achievements in Chromatography, The Ralph Adams Award in Bioanalytical Chemistry and several teaching awards. He has held several service posts and is presently Associate Editor of Analytical Chemistry and Chair of the Chemistry Department at University of Michigan.

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Cite this work

Researchers should cite this work as follows:

  • Robert Kennedy (2019), "Droplet Microfluidics for High-Throughput Chemical Analysis and Experimentation," https://nanohub.org/resources/30210.

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Location

WTHR 320, Purdue University, West Lafayette, IN

Tags

Droplet Microfluidics for High-Throughput Experimentation
  • Droplet Microfluidics for High-Throughput Experimentation 1. Droplet Microfluidics for High… 0
    00:00/00:00
  • Routes to Automation and High Throughput 2. Routes to Automation and High … 127.49416082749417
    00:00/00:00
  • Droplets in Microfluidic Systems 3. Droplets in Microfluidic Syste… 255.35535535535536
    00:00/00:00
  • Processing Nanoliter Samples Using Droplet Microfluidics 4. Processing Nanoliter Samples U… 319.019019019019
    00:00/00:00
  • High Throughput Reagent Addition in Droplet Format 5. High Throughput Reagent Additi… 496.59659659659661
    00:00/00:00
  • A Variable Nanoliter Pipetter 6. A Variable Nanoliter Pipetter 535.20186853520192
    00:00/00:00
  • Droplet Microfluidic Toolbox 7. Droplet Microfluidic Toolbox 586.38638638638645
    00:00/00:00
  • Analysis of Plug Contents: Direct ESI-MS 8. Analysis of Plug Contents: Dir… 657.62429095762434
    00:00/00:00
  • Direct ESI-MS of Segmented Flow 9. Direct ESI-MS of Segmented Flo… 699.4327660994328
    00:00/00:00
  • High Throughput MS Analysis of Plugs 10. High Throughput MS Analysis of… 732.59926593259934
    00:00/00:00
  • Low Flow Rates Beneficial for Complex Samples: GABA in 150 mM Ionic Strength Saline (aCSF) 11. Low Flow Rates Beneficial for … 758.692025358692
    00:00/00:00
  • Stable Long Term Analysis by nESI-MS 12. Stable Long Term Analysis by n… 851.951951951952
    00:00/00:00
  • Low Droplet Carry-Over During nESI-MS 13. Low Droplet Carry-Over During … 876.54320987654319
    00:00/00:00
  • High-Throughput at Low Volumes 14. High-Throughput at Low Volumes 945.2118785452119
    00:00/00:00
  • Electrophoretic Analysis of Droplets 15. Electrophoretic Analysis of Dr… 1000.266933600267
    00:00/00:00
  • Dual chip droplet extraction scheme 16. Dual chip droplet extraction s… 1076.3430096763429
    00:00/00:00
  • Electrophoresis from Droplets 17. Electrophoresis from Droplets 1114.4144144144145
    00:00/00:00
  • Droplet Microfluidics in Chemical Analysis 18. Droplet Microfluidics in Chemi… 1149.1491491491493
    00:00/00:00
  • 19. "Sensing" in the Living Brain:… 1191.257924591258
    00:00/00:00
  • In Vivo Measurements of Neurotransmitters 20. In Vivo Measurements of Neurot… 1240.573907240574
    00:00/00:00
  • Microdialysis Sampling for In Vivo Monitoring 21. Microdialysis Sampling for In … 1351.4848181514849
    00:00/00:00
  • Temporal Resolution in Microdialysis Sampling: Impact of Transfer Tubing 22. Temporal Resolution in Microdi… 1416.1161161161163
    00:00/00:00
  • No Temporal Distortion with Segmented Flows 23. No Temporal Distortion with Se… 1464.1641641641643
    00:00/00:00
  • On-Board Droplet Generator 24. On-Board Droplet Generator 1486.0860860860862
    00:00/00:00
  • Applications 25. Applications 1528.7287287287288
    00:00/00:00
  • Huntington's Disease 26. Huntington's Disease 1545.7457457457458
    00:00/00:00
  • Glutamate Neurotransmission 27. Glutamate Neurotransmission 1593.2265598932265
    00:00/00:00
  • Recovery of Normal Glu Uptake in HD Mice with Cef 28. Recovery of Normal Glu Uptake … 1655.4554554554554
    00:00/00:00
  • In Vivo Test – with MS Analysis 29. In Vivo Test – with MS Analy… 1745.9125792459126
    00:00/00:00
  • In Vivo Neurochemical Dynamics by Droplet MS 30. In Vivo Neurochemical Dynamics… 1823.7237237237239
    00:00/00:00
  • Droplet Microfluidics in Chemical Analysis 31. Droplet Microfluidics in Chemi… 1850.2168835502171
    00:00/00:00
  • High Throughput Screening (HTS) for Drug Discovery 32. High Throughput Screening (HTS… 1879.9466132799466
    00:00/00:00
  • Droplet Mass Spectrometry for HTS 33. Droplet Mass Spectrometry for … 1943.677010343677
    00:00/00:00
  • In-Droplet MS Assay (miniaturization) 34. In-Droplet MS Assay (miniaturi… 1973.4734734734736
    00:00/00:00
  • Can Droplet ESI-MS be Robust Enough to Screen Many Samples? 35. Can Droplet ESI-MS be Robust E… 2009.1758425091759
    00:00/00:00
  • Raw Data from Prestwick Library Screen 36. Raw Data from Prestwick Librar… 2053.9873206539874
    00:00/00:00
  • Electrophoresis for High Throughput Screening? 37. Electrophoresis for High Throu… 2097.6643309976644
    00:00/00:00
  • Electrophoresis for Screening: Fast Enzyme Assays 38. Electrophoresis for Screening:… 2170.4037370704036
    00:00/00:00
  • Large scale SIRT5 screening – 1280 Compounds 39. Large scale SIRT5 screening   2216.0160160160162
    00:00/00:00
  • Fast Sample Introduction: Caliper High Throughput 40. Fast Sample Introduction: Cali… 2265.5989322655992
    00:00/00:00
  • Comparison to LabChip System 41. Comparison to LabChip System 2327.7944611277944
    00:00/00:00
  • Protein Protein Interaction as Drug Targets 42. Protein Protein Interaction as… 2374.1408074741407
    00:00/00:00
  • Protein-Protein Interaction Assays and Screens 43. Protein-Protein Interaction As… 2453.1531531531532
    00:00/00:00
  • Affinity Probe CE (Noncompetitive Affinity Assay) 44. Affinity Probe CE (Noncompetit… 2471.4381047714382
    00:00/00:00
  • Hsp70 and Bag3 as Cancer Target 45. Hsp70 and Bag3 as Cancer Targe… 2520.286953620287
    00:00/00:00
  • Rapid Separation for Protein-protein interaction screening 46. Rapid Separation for Protein-p… 2573.23990657324
    00:00/00:00
  • Current Standard Screen: Flow Cytometry Protein Interaction Assay 47. Current Standard Screen: Flow … 2599.0323656990327
    00:00/00:00
  • Why is CE More Selective? 48. Why is CE More Selective? 2610.343677010344
    00:00/00:00
  • Conclusion 49. Conclusion 2653.7871204537873
    00:00/00:00
  • Enzyme Engineering For Catalysis 50. Enzyme Engineering For Catalys… 2675.508842175509
    00:00/00:00
  • Directed Evolution of Biocatalysts 51. Directed Evolution of Biocatal… 2759.75975975976
    00:00/00:00
  • Adapting Droplet Microfluidics to Directed Evolution 52. Adapting Droplet Microfluidics… 2855.7891224557893
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  • Summary 53. Summary 2945.8458458458458
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  • Acknowledgements 54. Acknowledgements 2992.1921921921921
    00:00/00:00
  • A Variable Nanoliter Pipetter 55. A Variable Nanoliter Pipetter 3444.8448448448448
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