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
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 Develop a miniaturized microfluidic-based cell culture apparatus for high throughput parallel drug apparatus for high throughput parallel drug
screening with on-chip monitoring of cell physiology.screening with on-chip monitoring of cell physiology.
Overall Project DefinitionOverall Project Definition
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
MotivationMotivation 128 well plate assay technique128 well plate assay technique Limitations: Limitations:
stationary state cell studystationary state cell study microliter reagent volumes microliter reagent volumes microliter scale studiesmicroliter scale studies
NanophysiometerNanophysiometer Nanoliter ScaleNanoliter Scale Real Time MonitoringReal Time Monitoring Decrease: Decrease: Reagents (if any!)Reagents (if any!)
ProcessingProcessing TimeTime
Project GoalsProject Goals Develop and Fabricate nanoliter sized cell Develop and Fabricate nanoliter sized cell
culture culture volumes with flow through perfusionvolumes with flow through perfusion Design on-chip peristaltic pumps for low Design on-chip peristaltic pumps for low
volume volume perfusion and drug administration. perfusion and drug administration. Design and incorporate thin film Design and incorporate thin film
microelectrodes microelectrodes in the inflow and outflow in the inflow and outflow structures for structures for differential electrochemical differential electrochemical monitoring of monitoring of various analytes such as pH, various analytes such as pH, oxygen, glucose oxygen, 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 NanoPhysiometermircofluidic control of the NanoPhysiometer
Nanoliter-sized Cell Nanoliter-sized Cell CultureCulture
** Nanophysiometer **** Nanophysiometer **
Accurate small-scale study of cell Accurate small-scale study of cell functionfunction
Flow Through PerfusionFlow Through PerfusionContinuous Media turn-overContinuous Media turn-over
Cell Cell FiltersFiltersSTATUS: Mask came in on Friday after 2 week delay!
500 um
300 um
Soft LithographySoft Lithography
STATUS: Made practice channels and will make master on Tuesday
Peristaltic PumpsPeristaltic Pumps
[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:
Have pneumatic controller and pneumatic mask are currently programming the Labview interface
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 cell lines are self sustaining and fluidics/pneumatics are performing
Physiometer Mask Physiometer Mask DesignDesign
Microfluidics
Pneumatics
Electrodes
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: Developing
pneumatic controller interface code
Labview Control PanelLabview Control Panel
IMAGE
SchematicSchematic
LabView
D/A Converter
Camera
Nanophysiometer
PneumaticController
Electrode
Optimize Cell Culture Optimize Cell Culture ConditionsConditions
Determine minimal Determine minimal flow rates for flow rates for maintaining vitalitymaintaining vitality
Allow for Allow for physiological physiological measurementsmeasurements Exp: pH differential as Exp: pH differential as
an indicator of an indicator of metabolic ratemetabolic rateSTATUS: This week we are
starting our own cell culture line of Fibroblasts from starter cells from Evgeni in ChemE Department.
Atmospheric Cell Culture Atmospheric Cell Culture ConditionsConditions
Maintain Maintain Temperature and Temperature and COCO22/O/O22 levels levels
PDMS gas PDMS gas permeablepermeable
Plexiglas Plexiglas enclosureenclosure
Heated Heated Microscope stageMicroscope stage
http://www.cyto.purdue.edu/flowcyt/educate/photos/confocal/images.htm
TimelineTimelineJANJAN FEBFEB MARMAR APRAPR
Literature & Patent Literature & Patent SearchSearch
Define Goals & Project Define Goals & Project ApproachApproach
Design MaskDesign Mask
Prototype/Device Prototype/Device FabricationFabrication
ProgrammingProgramming
OperationOperation
Assessment & EvaluationAssessment & Evaluation
Final Paper & Final Paper & PresentationPresentation
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 Nanodelivery, Inc.Nanodelivery, Inc.
