Instrumented Instrumented NanoPhysiometer for NanoPhysiometer for High Throughput Drug High Throughput Drug

ScreeningScreening

D. Michael Ackermann, Jon Payne, D. Michael Ackermann, Jon Payne,

Hilary Samples, James WellsHilary Samples, James Wells

Labview Front PanelLabview Front Panel

IMAGE

Big PictureBig PictureApplications:Applications:A Research ToolA Research Tool

Target Population:Target Population: Protoype of research tool Protoype of research tool Private research of BioMEMS group of Private research of BioMEMS group of

VUSE BME deptVUSE BME dept

Market Demand:Market Demand: Custom project for specific researchCustom project for specific research Future implications to broad marketFuture implications to broad market

High throughput screeningHigh throughput screeningPharmaceutical TestingPharmaceutical TestingToxicologyToxicology

MotivationMotivationLimited study of cell life, activity, and volumes Limited study of cell life, activity, and volumes

Previous methods:Previous methods:Single phase, stationary state Single phase, stationary state Microliter scale & volumesMicroliter scale & volumes

NanophysiometerNanophysiometer Nanoliter ScaleNanoliter Scale Real Time MonitoringReal Time Monitoring Decrease: Decrease: Reagents (if any!)Reagents (if any!)

ProcessingProcessingTimeTime

128 Well Plate Assays

Project GoalsProject Goals

Develop nanoliter sized cell culture volumeDevelop nanoliter sized cell culture volume On-chip pumps for low flow perfusion and On-chip pumps for low flow perfusion and

drug administration. drug administration. Thin film microelectrodes for monitoring of Thin film microelectrodes for monitoring of

various analytes such as pH, oxygen, various analytes such as pH, oxygen, glucose glucose and lactose in the media.and lactose in the media.

Optimize cell culture conditions to maintain Optimize cell culture conditions to maintain cell cell viability over long periods of time.viability over long periods of time.

Develop a Labview based user interface for Develop a Labview based user interface for mircofluidic control of the mircofluidic control of the

NanoPhysiometerNanoPhysiometer

Develop On-Chip Drug Delivery Systems

To Achieve Desirable Low Flow Profiles

Using Peristaltic Pumping

Providing Ideal Parameters for Cell

Viability

The NanoPhysiometerThe NanoPhysiometer

800 umGoals:

Physiometer Mask Physiometer Mask DesignDesign

Microfluidics

Pneumatics

Electrodes

800 um

Physiometer Design Physiometer Design ConcernsConcerns

Filter SizeFilter Size 3 um, 5 um, 8 um3 um, 5 um, 8 um

Channel Aspect Channel Aspect RatioRatio

Space between Space between fluidic and fluidic and pneumatic layerspneumatic layers

Peristaltic Pumps/ValvesPeristaltic Pumps/Valves

[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).

Fluidics Layer

Flexible PDMS Membrane (Valve)

STATUS:

Working! Currently just optimizing.

Electrochemical Electrochemical MonitoringMonitoring

Use integrated thin film Use integrated thin film microelectrodes to monitor microelectrodes to monitor physiological parametersphysiological parameters pH, glucose, etc.pH, glucose, etc.

Electrodes coated with a substrate Electrodes coated with a substrate specific oxidasespecific oxidase Catalyze reaction producing HCatalyze reaction producing H22OO22

HH22OO22 then detected then detectedSTATUS: Will be integrated once fluidics/pneumatics are performing optimally: hard to make!

Optimize Cell Culture Optimize Cell Culture ConditionsConditions

Determine minimal flow Determine minimal flow rates for maintaining rates for maintaining vitality & sufficient vitality & sufficient perfusionperfusion

Allow for physiological Allow for physiological measurementsmeasurements

Low flow for detectable pH Low flow for detectable pH and electrochemical and electrochemical differentialdifferentialSTATUS: We seeded and imaged fibroblast in devices of various sized filters for observation, testing of cell attachment , and minimal survival.

FIBROBLASTS

*3-8 m when spherical, (flat, dendrite-like when attached)

*1-2 day doubling time*Robust*Medium- antibiotics, vitamins, essential AA

Atmospheric Cell Culture Atmospheric Cell Culture ConditionsConditions

Cells demand optimal temperature and COCells demand optimal temperature and CO22/O/O22 levels levels PDMS is gas permeablePDMS is gas permeable

http://www.cyto.purdue.edu/flowcyt/educate/photos/confocal/images.htm

Plexiglas Plexiglas enclosureenclosure Contained, humidified Contained, humidified incubator environment incubator environment of 5% COof 5% CO22

Heated Heated Microscope stageMicroscope stage Maintains optimal Maintains optimal heated environment of heated environment of 37 C 37 C

LabView programmingLabView programming

User control of nanophysiometer User control of nanophysiometer system system Program Presets based on experimental Program Presets based on experimental

needsneeds Manual Control of Pumps and valvesManual Control of Pumps and valves

Measurements & Data acquisitionMeasurements & Data acquisition Show parameter measurementsShow parameter measurements

Time-Lapse Image CaptureTime-Lapse Image Capture Qualitative analysisQualitative analysisSTATUS: Nearly complete.

Labview Front PanelLabview Front Panel

IMAGE

SchematicSchematic

LabView

D/A Converter

Camera

Nanophysiometer

PneumaticController

Electrode

JANJAN FEBFEB MARMAR APRAPR

Literature & Patent Literature & Patent SearchSearch

Define Goals & Project Define Goals & Project ApproachApproach

Design MaskDesign Mask

Prototype/Device Prototype/Device FabricationFabrication

ProgrammingProgramming

Cell CultureCell Culture

OperationOperation

Assessment & EvaluationAssessment & Evaluation

Final Paper & PresentationFinal Paper & Presentation

BudgetBudget

Mask of device design- Mask of device design- $600/mask$600/mask

PDMS kit - $15PDMS kit - $15 Cell culture supplies- Cell culture supplies-

$300/month$300/month Tubing, wiring, etc.- ~$10 Tubing, wiring, etc.- ~$10 Electrodes- $500 Electrodes- $500 (owned by lab)(owned by lab)

ReferencesReferences Unger, Quake, et. al. Monolithic Microfabricated Unger, Quake, et. al. Monolithic Microfabricated

Valves and Pumps by Multilayer Soft Lithography.  Valves and Pumps by Multilayer Soft Lithography.  Science. Vol. 288.  April 7, 2000 Science. Vol. 288.  April 7, 2000

Ho, Chih-Ming. Fluidics – The Link Between Micro Ho, Chih-Ming. Fluidics – The Link Between Micro and Nano Sciences and Technologies. 0-7803-5998-and Nano Sciences and Technologies. 0-7803-5998-4/01. 2001 IEEE 4/01. 2001 IEEE

Arik, Zurn, et. al.  Design, Fabrication and Arik, Zurn, et. al.  Design, Fabrication and Experimental-Numerical Study of PZT Sensors. MSM Experimental-Numerical Study of PZT Sensors. MSM 2000.  Puerto Rico. 2000.  Puerto Rico.

Gonzalez, Moussa. Simulation of MEMS Piezoelectric Gonzalez, Moussa. Simulation of MEMS Piezoelectric Micropump for Biomedical Applications.  2002.  Algor Micropump for Biomedical Applications.  2002.  Algor Incorporated; Technical Document. Incorporated; Technical Document.

Bar-Cohen, Chang.  Piezoelectrically Actuated Bar-Cohen, Chang.  Piezoelectrically Actuated Miniature Peristaltic Pump.  March 2000.  Proceeding Miniature Peristaltic Pump.  March 2000.  Proceeding of 2000 SPIE Smart Structures and Materials of 2000 SPIE Smart Structures and Materials Symposium. No. 3992-103 Symposium. No. 3992-103

AcknowledgementsAcknowledgements

Dr. Franz BaudenbacherDr. Franz Baudenbacher David SchafferDavid Schaffer AndreasAndreas Nanodelivery, Inc.Nanodelivery, Inc.