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

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

Big Picture Applications High throughput screening Pharmaceutical Testing Cellular response to chemicals, toxins, etc. Contribute to Advancing area of  -Total Analysis Systems

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

Project Goals Develop and Fabricate nanoliter sized cell culture volumes with flow through perfusion 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 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. 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. 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 Develop a Labview based user interface for mircofluidic control of the NanoPhysiometer

Nanoliter-sized Cell Culture ** Nanophysiometer ** ** Nanophysiometer ** Accurate small-scale study of cell function Flow Through Perfusion Flow Through Perfusion Continuous Media turn-over Cell Filters Cell Filters STATUS: Culture chamber design selected and mask is sent out

Peristaltic Pumps [1] S.R. Quake and A. Scherer, "From Micro to Nano Fabrication with Soft Materials", Science 290: (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: (2000). Fluidics Layer Flexible PDMS Membrane (Valve) STATUS: Researched pump mechanisms and decided on pneumatic peristaltic pumps.

Electrochemical Monitoring Use integrated thin film microelectrodes to monitor physiological parameters Use integrated thin film microelectrodes to monitor physiological parameters pH, glucose, etc. pH, glucose, etc. Electrodes coated with a substrate specific oxidase Electrodes coated with a substrate specific oxidase Catalyze reaction producing H 2 O 2 Catalyze reaction producing H 2 O 2 H 2 O 2 then detected H 2 O 2 then detected STATUS: Will be integrated into nanophysiometer and used in testing culture conditions (begins next week!)

Physiometer Mask Design Microfluidics Pneumatics Electrodes

Optimize Cell Culture Conditions Determine minimal flow rates for maintaining vitality: Allow for physiological measurements Determine minimal flow rates for maintaining vitality: Allow for physiological measurements Exp: pH differential as an indicator of metabolic rate Exp: pH differential as an indicator of metabolic rate STATUS: Learned/practiced some cell culture technique outside of physiometer. Will begin optimizing flow condition for maintaining cells when nanophysiometer is built.

LabView programming User control of nanophysiometer system User control of nanophysiometer system Program Presets based on experimental needs Program Presets based on experimental needs Manual Control of Pumps and valves Manual Control of Pumps and valves Measurements & Data acquisition Measurements & Data acquisition Show parameter measurements Show parameter measurements Time-Lapse Image Capture Time-Lapse Image Capture Qualitative analysis Qualitative analysis STATUS: Design in progress

Labview Control Panel IMAGE

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