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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. Labview Front Panel. IMAGE. Big Picture Applications: A Research Tool. Target Population: Protoype of research tool

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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. Labview Front Panel IMAGE

  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 Limited study of cell life, activity, and volumes Previous methods: Single phase, stationary state Microliter scale & volumes • Nanophysiometer Nanoliter Scale Real Time Monitoring Decrease: Reagents (if any!) Processing Time 128 Well Plate Assays

  5. Project Goals • Develop nanoliter sized cell culture volume • On-chip pumps for low flow perfusion and drug administration. • Thin film microelectrodes for 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. The NanoPhysiometer Goals: 800 um Develop On-Chip Drug Delivery Systems To Achieve Desirable Low Flow Profiles Using Peristaltic Pumping Providing Ideal Parameters for Cell Viability

  7. Physiometer Mask Design Electrodes 800 um Microfluidics Pneumatics

  8. Physiometer Design Concerns • Filter Size • 3 um, 5 um, 8 um • Channel Aspect Ratio • Space between fluidic and pneumatic layers

  9. 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/Valves STATUS: Working! Currently just optimizing.

  10. 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 fluidics/pneumatics are performing optimally: hard to make!

  11. Optimize Cell Culture Conditions Determine minimal flow rates for maintaining vitality & sufficient perfusion • Allow for physiological measurements • Low flow for detectable pH and electrochemical differential FIBROBLASTS *3-8 mm when spherical, (flat, dendrite-like when attached) *1-2 day doubling time *Robust *Medium- antibiotics, vitamins, essential AA STATUS: We seeded and imaged fibroblast in devices of various sized filters for observation, testing of cell attachment, and minimal survival.

  12. Atmospheric Cell Culture Conditions • Cells demand optimal temperature and CO2/O2 levels • PDMS is gas permeable Plexiglas enclosure • Contained, humidified incubator environment of 5% CO2 Heated Microscope stage • Maintains optimal heated environment of 37 C http://www.cyto.purdue.edu/flowcyt/educate/photos/confocal/images.htm

  13. 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: Nearly complete.

  14. Labview Front Panel IMAGE

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

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

  17. 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

  18. Acknowledgements • Dr. Franz Baudenbacher • David Schaffer • Andreas • Nanodelivery, Inc.

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