harnessing microfluidics for research and development nathaniel c. cady asst. prof. nanobioscience...
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Harnessing Microfluidics for Research and Development
Nathaniel C. Cady
Asst. Prof. Nanobioscience
College of Nanoscale Science & Engineering
Outline
• Fluid dynamics (for non-majors)
• Building microfluidic devices
• Examples of research devices
Turbulent Flow
Laminar Flow
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
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”
Device Fabrication
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
Photolithography
Transfer Pattern
Develop Resist
Etch substrate
Remove Resist
Making a Microfluidic Device
Direct Indirect
Fabrication is relatively easy…
Practical Applications
Diagnostics
Integrated DNA Purification & Real-Time PCR
35mm
20 m
m
Microchip-based DNA Biosensor
GuSCN (lysis buffer)
EtOH (wash buffer) dH2O (elution buffer)
DNA-based Diagnostics
10 microns
Micropillars for DNA Purification
Integrated Control System
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
Practical Applications
Micro Printing & Patterning
30 microns
Biomolecular Printing
PEG Hydrogel
Glass
GoldSiO2
Signaling ?
With Dr. Bill Shain & Dr. Matt Hynd – Wadsworth Center, NYS Dept. of Health
Probing Neural Networks
Biomolecular Printing
Insert movie
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
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
Slow, difficult High acceleration / thermal exposure – potentially damaging to cells
Cellular Printing
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
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
Mouse MTLn3-GFP (diluted) printed on polystyrene
50 μm
Mammalian Cell Printing
Practical Applications
Cell Dynamics
O2 Input
Tumor Cell Input
Weir Structures
(Constrictions)
Collection Area
Output
1000µm
Cell
Weir Structures
(Constrictions)
Biomimetic Device for Tumor Cell Dissemination Studies
500µm
Device Filled with Dye
Fluid Flow Direction
Flow Cell Design
Units: (cm/sec)
500µm
Fluid velocity vectors
Fluid Dynamic Modeling
Flu
id F
low
100um
HEp3 Cells
(Human Epidermal Carcinoma 3)
Device Testing
Cells were smaller than anticipated – needed different weir spacing!
100um
Flu
id F
low
Rapid Prototyping of New Device
• 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
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