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Wheeler

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Wheeler

  1. 1. Microfluidic Technology for Assisted Reproduction Matthew B. Wheeler 1 and David J. Beebe 2 1 Department of Animal Sciences and Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign; 2 Department of Biomedical Engineering University of Wisconsin-Madison Acknowledgements Eric Walters and Sherrie Clark @ UIUC Henry “Gripp” Zeringue @ UW Lorraine Leibfried-Rutledge @ Bomed Kathy Haubert @ Vitae LLC Funding Sources: CFAR, NIH, UIUC, USDA, UW
  2. 2. Why micro?
  3. 3. Micro Bio Fluidics <ul><li>surface-to-volume ratio is 20,000 for 100 µm dia. </li></ul><ul><li>Reynolds number small </li></ul><ul><li>“ 2 phase” - cells, molecules, embryos </li></ul>For more see: E. M.Purcell, Life at Low Reynold’s Number, American Journal of Physics, 1977 Think at the scale
  4. 4. Low Reynolds Number Flow inertial viscous Laminar Brownian Motion Fluid Plugs Turbulence Improved manipulation? More in vivo -like?
  5. 5. Microfluidics Applications <ul><li>MIT technology review (2001) - “one of ten technologies that will change the world” </li></ul><ul><li>Markets </li></ul><ul><ul><li>Point of care diagnostics </li></ul></ul><ul><ul><li>Discovery/screening (not just drug) </li></ul></ul><ul><ul><li>DNA manipulation and processing </li></ul></ul><ul><ul><li>Analytical instruments </li></ul></ul><ul><ul><li>Drug delivery </li></ul></ul><ul><ul><li>Sensing </li></ul></ul><ul><ul><li>Assisted Reproduction </li></ul></ul><ul><ul><li>Bioproduction </li></ul></ul><ul><ul><li>Chemical engineering </li></ul></ul><ul><ul><li>Chemistry </li></ul></ul>Vitae
  6. 6. Microfluidic Large-Scale Integration Thorsen et al. Science 298: 580-584. A microfluidic chip, fabricated from silicone elastomer, that contains 2056 integrated microvalves in an area of one square inch. The chip is analogous to an electronic comparator and is an example of microfluidic large-scale integration. The complex plumbing in the chip allows 512 chambers to be mixed pairwise, with individual addressing and recovery of the results. [Photo: S. Maerkl]
  7. 7. Rationale Handling Steps in Conventional IVF Handling Steps in µchannel IVF Load oocyte/embryo Remove embryo Change medium and/or add Sperm, etc.
  8. 8. Hardware Traditional Inefficient & labor intensive µFluidic Automate procedures and improve efficiencies
  9. 9. Device Design <ul><li>Device Design </li></ul><ul><ul><li>Funnel - loading & unloading </li></ul></ul><ul><ul><li>Parking place - holding/placement </li></ul></ul><ul><ul><li>Channel - microenvironment </li></ul></ul><ul><ul><li>Wells - reservoir </li></ul></ul>
  10. 10. Micro Engineering <ul><li>Computer chip (IC) manufacturing </li></ul><ul><ul><li>Lithography, deposition, removal (etching) </li></ul></ul><ul><ul><li>Silicon, metals </li></ul></ul><ul><li>MEMS (MicroElectroMechanical Systems) </li></ul><ul><ul><li>IC methods to make mechanical things </li></ul></ul><ul><li>Micro Fluidics </li></ul><ul><ul><li>Borrowed from above to make small pipes </li></ul></ul><ul><ul><li>Largely glass & polymers </li></ul></ul><ul><ul><li>Caliper (Capillary Electrophoresis) </li></ul></ul><ul><ul><li>Micro arrays (Nanogen, Affymetrix) </li></ul></ul>
  11. 11. Basic Logistics <ul><li>Loading/unloading </li></ul><ul><li>Transport (no cumulus) </li></ul><ul><li>Transport (with cumulus) </li></ul>
  12. 12. Chemical Manipulation <ul><ul><li>“ Parking place” </li></ul></ul>Zona Removal Device 200  m 200  m 7.5  m Before After
  13. 13. IVP (mice & pigs) <ul><li>General conditions </li></ul><ul><ul><li>Static (“no flow”) </li></ul></ul><ul><ul><li>Straight channels (250µm high x 1000 µm wide) </li></ul></ul><ul><li>Demonstrated </li></ul><ul><ul><li>IVM, IVF, EC </li></ul></ul>
  14. 14. Mouse Culture (Development) Microchannel Control Improved efficiencies % Blastocyst B6SJL/F1 x ICR - less vigorous
  15. 15. Mice, Pigs and Cows
  16. 16. What’s really going on? Micro fluidics Reduced “effective” volume Environmental control Micro environment
  17. 17. Micro Fluidics <ul><li>Effective volume </li></ul><ul><ul><li>~50 µl vs. ~ 50 nl </li></ul></ul><ul><li>Environmental control </li></ul><ul><ul><li>Rapid, precise stimulus application </li></ul></ul><ul><ul><li>Gradual, gentle media changes </li></ul></ul><ul><li>Micro environment </li></ul><ul><ul><li>Diffusion governs transport </li></ul></ul><ul><ul><li>Good & bad stuff hangs around </li></ul></ul>
  18. 18. <ul><li>1). Food and water supply monitoring: </li></ul><ul><li>-presence of residues, trace chemicals, </li></ul><ul><li> antibiotics, pathogens, toxins); </li></ul><ul><li>- integrated, rapid DNA sequencing to identify </li></ul><ul><li> genetic variation and GMO’s; </li></ul><ul><li>-integrity of food during transportation and storage </li></ul><ul><li>Animals health monitoring: </li></ul><ul><li>-developmental biology; </li></ul><ul><li>-presence of residues, antibiotics, pathogens, toxins; </li></ul><ul><li>-bio-sensors </li></ul><ul><li>Environment monitoring: </li></ul><ul><li>-land, water and air pollution; </li></ul><ul><li>-remote/distributed sensing </li></ul>Opportunities for Nanotechnology in Agriculture and Food Systems Research
  19. 19. 1). Develop agriculture and food systems related microfluidic devices that feature integrated operations, simple reliable components and low costs. 2). Integrate microfluidic devices flawlessly into a wireless-ID network. 3). Others ? ? ? ? Objectives of a National Research Program
  20. 20. 1). The users are producers, processors and consumers. -applications must be simple, reliable and highly accurate 2). The “environment” is dirty. -samples need some processing (filtration, purification, etc.) 3). Agriculture and Food Systems are highly integrated. -applications need to be networked and results integrated 4). Single sensor applications are likely not inadequate. -fields, herds, flocks, elevators, trains, trucks, processors, manufacturers etc. are widespread How is Agriculture Different? (or how will USDA’s focus differ from DOD, NSF, NIH or NASA) Outcomes and impacts should address these issues!
  21. 21. Research Budget Estimate <ul><li>1). Biochemical/Genetic/Residue detection </li></ul><ul><li>-$5 million/year </li></ul><ul><li>2). Systems for high-throughput drug/cell </li></ul><ul><li>screening and biosensor applications </li></ul><ul><li>-$5 million/year </li></ul><ul><li>3). Automated/integrated networks </li></ul><ul><ul><ul><ul><ul><li>-$3 million/year </li></ul></ul></ul></ul></ul><ul><li>4). Application testing </li></ul><ul><li>-$3 million/year </li></ul>

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