instrumented nanophysiometer for high throughput drug screening
DESCRIPTION
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 - PowerPoint PPT PresentationTRANSCRIPT
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.