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Instrumented NanoPhysiometer for High Throughput Drug Screening

Instrumented NanoPhysiometer for High Throughput Drug Screening. D. Michael Ackermann, Jon Payne, Hilary Samples, James Wells. Overall Project Definition.

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Instrumented NanoPhysiometer for High Throughput Drug Screening

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  1. Instrumented NanoPhysiometer for High Throughput Drug Screening D. Michael Ackermann, Jon Payne, Hilary Samples, James Wells

  2. Overall Project Definition Develop a miniaturized microfluidic-based cell culture apparatus for high throughput parallel drug screening with on-chip monitoring of cell physiology. Develop On-Board Drug Delivery Systems To Achieve Desirable Low Flow Profiles Using Peristaltic Pumping Providing Ideal Parameters for Cell Viability

  3. Big PictureApplications:A Research Tool • Target Population: • Protoype of research tool • Private research of BioMEMS group of VUSE BME dept • Market Demand: • Custom project for specific research • Future implications to broad market High throughput screening Pharmaceutical Testing Toxicology

  4. Motivation • 128 well plate assay technique Limitations: stationary state cell study microliter reagent volumes microliter scale studies • Nanophysiometer Nanoliter Scale Real Time Monitoring Decrease: Reagents (if any!) Processing Time

  5. Project Goals • Develop and Fabricate nanoliter sized cell culture volumes with flow through perfusion • Design on-chip peristaltic pumps for low volume perfusion and drug administration. • Design and incorporate thin film microelectrodes in the inflow and outflow structures for differential electrochemical monitoring of various analytes such as pH, oxygen, glucose and lactose in the media. • Optimize cell culture conditions to maintain cell viability over long periods of time. • Develop a Labview based user interface for mircofluidic control of the NanoPhysiometer

  6. Nanoliter-sized Cell Culture • ** Nanophysiometer ** Accurate small-scale study of cell function • Flow Through Perfusion Continuous Media turn-over • Cell Filters 300 um 500 um STATUS: Mask came in on Friday after 2 week delay!

  7. Soft Lithography STATUS: Made practice channels and will make master on Tuesday

  8. Fluidics Layer Flexible PDMS Membrane (Valve) [1] S.R. Quake and A. Scherer, "From Micro to Nano Fabrication with Soft Materials", Science 290: 1536-40 (2000). [2] M.A. Unger, H.-P. Chou, T. Thorsen, A. Scherer, and S.R. Quake, "Monolithic Microfabricated Valves and Pumps by Multilayer Soft Lithography", Science 288: 113-116 (2000). Peristaltic Pumps STATUS: Have pneumatic controller and pneumatic mask are currently programming the Labview interface

  9. Electrochemical Monitoring • Use integrated thin film microelectrodes to monitor physiological parameters • pH, glucose, etc. • Electrodes coated with a substrate specific oxidase • Catalyze reaction producing H2O2 • H2O2 then detected STATUS: Will be integrated once cell lines are self sustaining and fluidics/pneumatics are performing

  10. Physiometer Mask Design Electrodes Microfluidics Pneumatics

  11. LabView programming • User control of nanophysiometer system • Program Presets based on experimental needs • Manual Control of Pumps and valves • Measurements & Data acquisition • Show parameter measurements • Time-Lapse Image Capture • Qualitative analysis STATUS: Developing pneumatic controller interface code

  12. Labview Control Panel IMAGE

  13. Schematic Camera LabView Nanophysiometer Electrode Pneumatic Controller D/A Converter

  14. Optimize Cell Culture Conditions • Determine minimal flow rates for maintaining vitality • Allow for physiological measurements • Exp: pH differential as an indicator of metabolic rate STATUS: This week we are starting our own cell culture line of Fibroblasts from starter cells from Evgeni in ChemE Department.

  15. Atmospheric Cell Culture Conditions • Maintain Temperature and CO2/O2 levels • PDMS gas permeable • Plexiglas enclosure • Heated Microscope stage http://www.cyto.purdue.edu/flowcyt/educate/photos/confocal/images.htm

  16. Timeline

  17. Budget • Mask of device design- $600/mask • PDMS kit - $15 • Cell culture supplies- $300/month • Tubing, wiring, etc.- ~$10 • Electrodes- $500 (owned by lab)

  18. References • Unger, Quake, et. al. Monolithic Microfabricated Valves and Pumps by Multilayer Soft Lithography.  Science. Vol. 288.  April 7, 2000 • Ho, Chih-Ming. Fluidics – The Link Between Micro and Nano Sciences and Technologies. 0-7803-5998-4/01. 2001 IEEE • Arik, Zurn, et. al.  Design, Fabrication and Experimental-Numerical Study of PZT Sensors. MSM 2000.  Puerto Rico. • Gonzalez, Moussa. Simulation of MEMS Piezoelectric Micropump for Biomedical Applications.  2002.  Algor Incorporated; Technical Document. • Bar-Cohen, Chang.  Piezoelectrically Actuated Miniature Peristaltic Pump.  March 2000.  Proceeding of 2000 SPIE Smart Structures and Materials Symposium. No. 3992-103

  19. Acknowledgements • Dr. Franz Baudenbacher • David Schaffer • Nanodelivery, Inc.

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