intro to lab 3: modeling a microvascular network on a chip
DESCRIPTION
BME1310 17.3.2011 Erik Zavrel. Intro to Lab 3: Modeling a Microvascular Network on a Chip. Lab Objective. To characterize a simple model of microvascular system using a microfluidic device AND To introduce you to microfluidics - PowerPoint PPT PresentationTRANSCRIPT
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Intro to Lab 3: Modeling a Microvascular Network on a Chip
BME131017.3.2011Erik Zavrel
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Lab Objective To characterize a simple model of microvascular
system using a microfluidic device AND To introduce you to microfluidics
Measure flow velocities within model network before and after a blockage (mimicking an occlusion of a blood vessel or clot like during a stroke) and track the redistribution of flow when a channel is occluded.
Lab will be followed by a homework where you will theoretically calculate the pressure and velocity in each channel before and after blockage using MATLAB. You will then compare your calculated pressure and velocities with your measured velocities.
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What is Microfluidics: What’s in a name? Infer: manipulation of minute quantities of fluids (L and G) …
inside devices made using advanced microfab tools (CNF). Applies ultra-precise fab technology to conventionally “messy” fields like
biology (dishes, plates) and chemistry (vats, reaction vessels) New field – emerged early 90s: Andreas Manz, George Whitesides
Why?▪ Unique physics at microscale permits novel creations:
▪ Laminar flow▪ High SA/V ratio – surface tension (droplet) >> gravity (sedimentation)
▪ Reduces amounts of reagents needed▪ Permits more orderly, systematic approach to bio-related problems, reduces physical
effort: drug discovery, cytotoxicity assays, protein crystallization (for x-ray crystallography)
▪ Disposable, parallel operation, increased reliability
Applications:▪ Biomimesis, diagnostic devices, biosensors, cell sorting, enrichment, storing
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Examples of Microfluidics
LSI Body on chip Concentration gradient
Sperm sorter for IVF
Traction force microscopy
Portable medical
diagnostics
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Your Microfluidic Device
5 mm square 5 mm square
200 m x 5000 m
200 m x 5000 m
50 m x 500 m100 m x 5000 mHeight = 100 m
200 m x 4300 m
200 m x 8600 m
Mimics this
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Procedure: Before Clot
PDMS devices made for you Fill syringe with bead (10um dia.) solution Connect needle to syringe Connect tubing to needle Place syringe in syringe pump Flow @ 50uL/min to flush out bubbles Reduce flow rate to 1-5uL/min Capture series of images to track beads
in each channel of device V = d/t
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Procedure: Clot
Disconnect device from pump Using empty syringe, introduce air to
dry device Punch hole in desired channel with
punch Remove plug of PDMS Inject sealant to block channel Cure @ 60C for 10min Repeat process: track beads in each
channel
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How Many Channels?
5 mm square 5 mm square
200 m x 5000 m
200 m x 5000 m
50 m x 500 m100 m x 5000 mHeight = 100 m
200 m x 4300 m
200 m x 8600 m
12
3
4
56
7
8 9
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Results and Data Analysis Expected
Average velocity for each channel before blockage Average velocity for each channel after blockage Prediction of flow pattern after channel blockage Bead Motion Analysis:
Want to track at least 10 beads in each channel Track beads in each channel over several frames Not interested in instantaneous velocity of beads
(changes from frame to frame) Interested in average velocity (total distance traveled /
time observed) Bin data to generate histogram (data partitioned into
intervals and frequency of occurrence plotted)
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Note on Bead Motion
In pressure-driven flow, velocity profile of fluid is parabolic:
Velocity is maximum in center of channel
Velocity is minimum near walls of channel
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Report Guidelines Title Page Abstract Introduction Materials and Methods Results Discussion Questions
Place raw data in appendix Place processed, presentable data in body of report
Cite at least 3 sources posted on Blackboard
MATLAB session to be held in Carpenter tentatively set for Fri April 15th ImageJ covered in lab
Due Thursday 14th of April in class
Bring USB to save large data files!
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Analysis: Circuit Analog In analyzing a microfluidic device, it is useful to make an analogy to an electrical
circuit: Pressure → Voltage Flow Rate → Current Hydraulic Resistance → Electrical Resistance Volumetric flow rate = average linear velocity * cross-sectional area of channel Ohm’s Law:
V = IR Pressure = Volumetric Flow Rate * Channel Resistance
Ground = reference potential → same pressure KCL:
Conservation of charge → conservation of mass:▪ Flow rate into a node MUST equal flow rate out of that node▪ Nothing collects at node i1 → ←i2
i3 ↓
Convention: Define flow into node as + and flow out of node as –I1 + i2 – i3 = 013 = i1+i2
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Circuit Analysis Resistances are either in series or
parallelR 1
1k
R 2
2k
R 3
3k
ba
R 11k
R 22k
R 35k
b
a
RT = R1+R2+R3 = 1000+2000+3000 = 6000 ohm
Sum is ALWAYS greater than any single resistanceRT = 1 / (1/R1 + 1/R2 + 1/R3) =
1/ (1/1000 + 1/2000 + 1/3000) = 545 ohm
Sum is ALWAYS less than any single resistance
Special Case: 2 in parallel:RT = R1*R2 / (R1+R2)
Series:
Parallel:
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Example: Resistance Network
R 1
1k
R 2
5k
R 3
250
R 5
1500
R 6
250 R 7400
b
a
R 4
250
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1. R1 and R2 in series = 6k
6k
R 3
250
R 5
1500
R 6
250 R 7400
b
a
R 4
250
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2. R3 and R4 in series = 500 ohm
6k
500
R 5
1500
R 6
250 R 7400
a
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3. 500 ohm in parallel with 1500 ohm = 375 ohm
6k 375
R 5
250 R 6400
b
a
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R T
7025
a b
Remaining resistors in series = 6000 + 375 + 250 + 400 = 7025 ohm
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Pressure Drop Along ChannelEquation valid only if w >>h
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Pressure Drop Along Channel
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Fabrication: How Are Microfluidic Devices Made? Pattern is defined on surface (2D)
and then transferred into vertical plane
Process flow – depicted as a cross-sectional view, each step showing execution of one step (addition of layer, exposure, removal of layer, etc.)
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Process Flow: Soft-Lithography
1) Spin SU-8
Si
SU-8
Mask
2) Expose
3) Develop
4) Apply Anti-stick monolayer
6) Plasma treat + bond
glass
2) Expose
mask
5) Cast PDMS
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