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Harnessing Microfluidics for Research and Development
Nathaniel C. Cady
Asst. Prof. Nanobioscience
College of Nanoscale Science & Engineering
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Outline
• Fluid dynamics (for non-majors)
• Building microfluidic devices
• Examples of research devices
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Turbulent Flow
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Laminar Flow
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Re = (density) x (velocity) x (diameter) (viscosity)
If Re = 3000 or higher = turbulent flow
If 2000-3000 = transitional flow
If less than 2000 = laminar flow
2300 = transition point
Reynolds Number
Flow regime is predictable!
d
v
Microfluidic devices capitalize on small channel sizes to control flow regime
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Advantages of Microfluidic Devices
• Well-controlled fluid dynamics
•Diffusion-limited mixing
•Controllable fluid interactions
• Small fluid volume
•Less sample and reagent needed
•More samples per unit area (multiplexing)
Microfluidics = “Lab-on-a-chip”
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Device Fabrication
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Fabrication of Microfluidic Devices
• Fabrication schemes range from simple to highly complex
• Primarily rely on micro / nanofabrication techniques
• Lithography (photo-, electron beam, imprint)
• Etching or molding of 3-D channels
• “Capping” or enclosure of channels
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Photolithography
Transfer Pattern
Develop Resist
Etch substrate
Remove Resist
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Making a Microfluidic Device
Direct Indirect
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Fabrication is relatively easy…
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Practical Applications
Diagnostics
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Integrated DNA Purification & Real-Time PCR
35mm
20 m
m
Microchip-based DNA Biosensor
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GuSCN (lysis buffer)
EtOH (wash buffer) dH2O (elution buffer)
DNA-based Diagnostics
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10 microns
Micropillars for DNA Purification
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Integrated Control System
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Category Organism / Target DNA Purification Real-Time Detection Detection Limit
Bacteria Salmonella typhimurium Yes Yes 10 cells
Bacillus anthracis (Sterne) Yes Yes 40 cells
Listeria monocytogenes Yes Yes 100 cells
Staphylococcus aureus Yes Yes --
Escherichia coli Yes Yes --
Bacillus globigii (subtilis ) Yes Yes --
Phage Lambda Yes Yes --
Parasites Leishmania donovani Yes Yes --
Human CYP3A56 (SNP) Yes Yes --
ABCA1 (SNP) Yes Yes --
Amelogenin (SNP, gender) Yes Yes --
Cady et. al. (2005) Sensors & Actuators B. 107(1): 332-341
Cady et. al. (2003) Biosensors & Bioelectronics. 19: 59-66
Detection Results
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Practical Applications
Micro Printing & Patterning
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30 microns
Biomolecular Printing
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PEG Hydrogel
Glass
GoldSiO2
Signaling ?
With Dr. Bill Shain & Dr. Matt Hynd – Wadsworth Center, NYS Dept. of Health
Probing Neural Networks
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Biomolecular Printing
Insert movie
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Microelectrode Array (MEA) Hydrogel-coated MEA patterned with the
laminin peptide, biotin-IKVAV. The laminin peptide biotin-IKVAV was printed onto using the automated NanoEnabler bioprinter. Printed peptide was arranged in a pattern consisting of orthogonal 2 mm-wide lines connecting 10 mm diameter node.
200um
• Microelectrode arrays (MEAs) coated with PEG-based hydrogel
• NeN used to pattern hydrogel with FITC-labeled bioactive peptides
• Successful printing of both spots and lines
Courtesy of: Matthew Hynd, PhD – NYS DOH
Printed Guidance for Neural Networks
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Patterned neuronal network at 2 weeks in vitro. Primary hippocampal neurons were plated onto patterned arrays at a density of 400 cells/mm2.
Scanning electron microscope image of patterned neural network.
200um
• Printed MEAs seeded with primary hippocampal neurons
• Cells proliferated on the arrays and formed neural network on MEAs
• Results were comparable to studies using microcontact printing methods (Hynd, et. al., J. Neuroscience Methods, 2006)
Courtesy of: Matthew Hynd, PhD – NYS DOH
Neural Networks
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Slow, difficult High acceleration / thermal exposure – potentially damaging to cells
Cellular Printing
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Fluid Reservoir
Channel
Printing Tip
Polymeric Surface Patterning Tool
• Developed at CNSE, UAlbany (Cady Lab)
• Designed to enable live cell printing directly onto solid surfaces
• Larger channels and cantilever allow for whole cells to be printed
30 microns
BioForce Silicon-based SPT
Polymeric SPT
Direct Cell Printing
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E. coli pET28A-GFP on polystyrene
100um100um
20 μm 20 μm 20 μm
Bacterial Cell Printing
E. coli pET28A-GFP TSA Plate (12 hr)
100um
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Mouse MTLn3-GFP (diluted) printed on polystyrene
50 μm
Mammalian Cell Printing
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Practical Applications
Cell Dynamics
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O2 Input
Tumor Cell Input
Weir Structures
(Constrictions)
Collection Area
Output
1000µm
Cell
Weir Structures
(Constrictions)
Biomimetic Device for Tumor Cell Dissemination Studies
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500µm
Device Filled with Dye
Fluid Flow Direction
Flow Cell Design
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Units: (cm/sec)
500µm
Fluid velocity vectors
Fluid Dynamic Modeling
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Flu
id F
low
100um
HEp3 Cells
(Human Epidermal Carcinoma 3)
Device Testing
Cells were smaller than anticipated – needed different weir spacing!
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100um
Flu
id F
low
Rapid Prototyping of New Device
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• Microfluidic devices reduce sample volume and offer unique fluid dynamic environments
• Novel fluid dynamics can affect reaction rates, diffusion, biological processes
• Practical applications (like patterning) can be accomplised using microfluidics
• Novel fluid environments can be used for biomimetic studies
Summary
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Acknowledgements University at Albany Mt. Sinai
Dr. Robert Geer Dr. Julio Aguirre-Ghiso
Dr. Magnus Bergkvist
Dr. Alain Kaloyeros
Research Support
UAlbany Startup Funds
UAlbany FRAP A&B Awards
BioForce Instruments
CNSE / CAS Challenge Grant