Advanced Manufacturing of Integrated
Labs-on-a-Chip for Ubiquitous Diagnostics
Gisela Lin, Ph.D. MEPTEC, San Jose, CA May 22, 2013
Micro/Nano Fluidics Fundamentals Focus (MF3) Center
What is Micro/Nano Fluidics?
Initially borrowed integrated circuit fabrication techniques to make mechanical as well as electrical components on a single chip.
Small size channels, wells, pumps, valves on the order of 1μm – 1mm
Even smaller functionalized surfaces, quantum dots, etc. (nanoscale)
Applications: Genetic analysis, proteomics, diagnostics, biosensing, bio-
imaging, drug delivery, cellular manipulation, microsurgical tools…
Disposable
“Lab-on-a-Chip”
Drawing and photos courtesy of Caliper Technologies, Inc.
Early Silicon-based Labs-on-a-chip
Mixing
chamber
Bubble pump
valve
Aluminum interconnects
Polysilicon heater
Fluid
channel
J. Evans, D. Liepmann, and A. Pisano (BSAC/UCB)
750m
Silicon/glass common (ink jet print heads, valves, pumps, etc.).
25µm SOI wafer, DRIE etch of silicon, bond clear glass cover-plate on top
Need off-chip pumping, electrical connections. Cost/device relatively high.
Early Polymer-based Labs-on-a-chip
PDMS = poly-dimethylsiloxane (silicone rubber)
Usually silicon-based mold, cast & cure PDMS replica. Scale up difficult.
Need external supporting equipment, need trained personnel to operate
Photo courtesy of Fluidigm, Inc.
3D network can contain as many as 600
valves controlled by 12 fluid control lines.
World-to-chip interface can get
complicated – need off-chip controllers
and pressure/vacuum sources.
Samples pipetted in by hand.
Lab-on-a-Chip vs. Chip in a Lab!
chip
Chips themselves have limited functionality, made one by one or in small batches via
custom processes in academic labs.
Very few commercial labs-on-a-chip (LOCs), and those that exist require supporting
equipment (benchtop or handheld reader, etc.) and trained personnel to operate.
MF3 Center formed to addresses these challenges.
Micro/nano Fluidics Fundamentals Focus Center
GOAL: Bridge the gap between academia and industry
– MF3 is a focused community that performs fundamental micro/nano fluidic research to develop standardized integration processes & device technology expedition of micro/nano
fluidic commercialization.
– Initiated in October 2006, headquartered at UCI, total funding = $12.5M over 6 years.
– Funded by DARPA and Corporate Members, 1 of 7 DARPA S&T centers
– 20 faculty at 12 universities + 7 companies + 2 government labs working together
Development of New
Microfluidic Solutions
Publications
Prototype platforms,
Graduates skilled in microfluidic
technology
Funds,
applications
Students,
expertise
Products
for national
interests
Industry Academia
Funds,
applications
Revenue
Funds for
educating &
training
Government
MF3 Center Goals
Our Mission: Create a focused community that performs
fundamental micro/nano fluidic (MF) research to develop
standardized MF integration processes and device technology
that results in the expedition of MF commercialization.
Work with corporate
partners to adapt,
consolidate, integrate
standardize
Barriers: Many different
technologies, fabrication
processes. Need to be
more application-driven.
Commercialization,
manufacturing,
volume production
Our E-Health Vision
Create “Ubiquitous Diagnostics”
– Integrated, low-cost, simple, labs-on-a-chip that can rapidly perform assessment anywhere and everywhere
– Environment, agriculture, food and water supplies, and ultimately for human health and safety.
– Labs-on-a-chip produced via high volume manufacturing processes
Interface chips with existing communications
infrastructure for data handling
– Cellular phones – Portable computers – Cloud computing – Social media
Manufacturing Processes
PCB Microfluidics USB Microfluidics Mobile device = power
source & data transmission
PRINTED CIRCUIT BOARD TECHNOLOGY
Prototyping, small to
mid-scale injection molding Contract manufacturing,
high volume molding
University laboratory
design/prototype
INJECTION MOLDING
Roll-to-roll Manufacturing Processes
Large scale roll-to-roll paper printing
Roll-to-roll Atomic Layer Deposition (Image courtesy of Beneq)
Large Roll Flex Circuit Manufacturing (image courtesy of Automated Assembly Corp.)
Roll-to-roll hot embossing (Image courtesy of VTT)
Microfluidics (hot embossed plastic)
Assay (printed, die-cut paper)
Electronics (printed/laminated
flexible circuit)
Integrated, multiple
layer, multi-functional,
low-cost microfluidic
platform
Combine manufacturing processes to create fully capable
low-cost diagnostic labs-on-a-chip
Lab-on-a-Chip Manufacturing Vision
Ubiquitous Diagnostics for E-Health
Utilize existing
communications
Sterilize and package the chips just
like Band-Aids!
A different chip for different assays
microfluidic
platform
Ubiquitous Diagnostics for E-Health
Satellite Integrated, low-cost, simple
labs-on-a-chip that can
quickly perform assessment
anywhere and everywhere.
Epidemiology
Agriculture
Military
Food/water supply
Hospitals
Home healthcare
Environment
Recent progress towards
ubiquitous diagnostics at the
MF3 Center
Particle Separation: Inertial Microfluidics
Particles experience
Dean drag force along
with inertial force in a
curved channel (Dean
flow = secondary
rotational flow field
perpendicular to flow
direction, which
produces drag force).
Particles occupy a
single equilibrium
position near the inner
channel wall,
depending on ratio of
inertial lift to Dean
drag.
Illustration courtesy of M. Toner et. Al. , New Journal of Physics, 2009.
Ian Papautsky (U. Cincinnati)
PDMS device separating 10µm (purple), 15µm (green), and 20µm (red)
diameter particles.
R2R Device for Blood Cell Sorting
Spiral channels for sorting blood cells
Fabricated via R2R hot embossing of
PMMA film
Mixture of cells
in whole blood
Plasma,
platelets
Erythrocytes
Leukocytes
Ian Papautsky (U. Cincinnati)
Lateral Cavity Acoustic Transducer
Air
Liquid Air-liquid
interface
Flow
direction
PUMP CONFIGURATION:
Side channel oriented 15º to main
channel
Acoustic streaming produces net force
on bulk fluid, pushing it forward.
Lateral Cavity Acoustic Transducer (LCAT): Dead end side channel traps air.
Vibrating the air/liquid interface via PZT disk results in acoustic streaming.
Single mask design facilitates easy integration with other MF components.
Abe Lee (UC Irvine)
LCAT for Blood Separation & Cell Lysing
20 sec lyse 2 min lyse
Inlet
Outlet
FLOW
Hgb Absorbance vs.
Lysing time
Blood cells Plasma
Blood
cells get
trapped in
vortices
Abe Lee (UC Irvine)
LCAT – Versatile microfluidic platform
Abe Lee (UC Irvine)
Portable Microfluidic Systems
An iPhone controlled microfluidic pumping manifold is demonstrated and is
one of the efforts towards universal portable MF platforms.
Next-gen LCAT devices implemented in R2R hot embossed plastic films.
Abe Lee (UC Irvine)
Paper-based microfluidics – Early devices
Sample (blood, urine, water, etc.)
Paper separates particulates out!
Photoresist
(hydrophobic)
Exposed
paper
1mm channels
(hydrophillic)
Protein & glucose assays
Small, light-weight, low-power, easy-to-use, field-deployable solider health and
environmental diagnostic devices that use existing communications infrastructure.
Third-world countries, remote locations for health and environmental monitoring.
Less than 1¢ per assay. Sample-to-answer in ~ 25min.
1.5 cm
Scan then upload to internet OR
photograph then transmit via phone
0
10
20
30
40
50
60
0 10 20 30 40 50 60 70
Mea
n In
ten
sity
[BSA], M
Calibration Curve for BSA
George Whitesides (Harvard University)
Multilevel Paper Microfluidics
Sample distribution,
sample sorting
FRONT BACK
1cm
1cm
OVERPASS
VIA
Added functionality – multilevel devices:
Stack layers of
patterned paper and
double-sided tape.
Fluids move laterally in
channels patterned in
the paper.
Fluids move vertically
through holes
patterned in the tape.
3D Diagnostics
BEGIN END
sample control Glucose
assay Protein
assay George Whitesides (Harvard University)
Wax Printing of Paper LOCs
Simple process using laser printing of wax-based ink
Melt wax to create complete fluidic barrier
Idea to device in minutes!
George Whitesides (Harvard University)
0.5cm
Inexpensive Large Scale Printing
5 cents/page ≈ 0.1 cents/device
Price/device decreases with scale up (i.e. roll-to-roll printing)
George Whitesides (Harvard University)
Paper-based liver function test
Analytes are ALP = alkaline phosphatase, AST = aspartate aminotransferase, and
total serum protein.
Device has 4 integrated components: (i) top plastic sheet, (ii) filter membrane to
separate blood cells from plasma – sample prep, (iii) patterned paper chip containing
the reagents necessary for analysis, (iv) bottom plastic sheet .
Compare
colorimetric output to
calibration curves
Analytical Chemistry
February 2012
Microfluidic Digital Logic
Achieve monolithic integration by using microfluidic
circuits to implement control logic.
Get rid of complex connections to
off-chip controllers!
Normally closed valve analogous to NMOS
Fundamental Boolean logic gates Elliot Hui (UC Irvine)
Semi-autonomous Liquid Handling
State Selectors
Vacuum
Supply
Peristaltic Pump Control
Boolean
Logic
Block
Resistor Network
Peristaltic Pump Control
Ring
Mixer
Pumps
Device contains oscillators, clocks, counters, pumps, and a 2-bit finite state
machine capable of cycling through 4 states (00, 01, 10, 11).
Chip = 2 sheets of machined plastic or etched glass + 1 sheet elastomer
Entire device (control + fluid handling) driven off a single vacuum source.
Elliot Hui (UC Irvine)
Semi-autonomous Liquid Handling
Elliot Hui (UC Irvine)
Portable Vacuum Sources - Options
A bicycle pump can work for hours while mouth suction and syringe pull
can achieve useable vacuum for a few minutes.
Elliot Hui (UC Irvine)
1 mm
Polyurethane:
2x 15 µL wells
Channel = 3.7 cm x 300 µm x70 µm
PCB Planarization
Fluidics
Sealant
Thermal Component Thermal Component
100 µm
PCBs are manufacturable and enable easy integration with standard electronics.
Thermal component contains 4 resistive heaters and a temperature sensor.
Integrated on-chip heaters, temperature sensors , electrical leads to achieve thermal
cell lysis, convective mixing, and nucleic acid extraction on a single platform.
PCB-based Lab-on-a-Chip
Mark Bachman (UC Irvine)
Heater, temp sensors
inside well
DNA Isolation via Isotachophoresis (ITP)
Collection
outlet
Inlet
ITP = species separation by
mobility under applied electric
field.
TE = trailing electrolyte, LE =
leading electrolyte
Demonstrated with cell culture,
urine, pathogens in blood, and
host blood nucleic acid (NA),
DNA, and RNA. Analytical Chemistry October 2012
Mark Bachman (UC Irvine) & Juan Santiago (Stanford)
Channel
Demo: Malaria-Infected Whole Blood
1 mm
Threshold qPCR Cycle vs. Parasite Concentration
Inlet well heating stirs, lyses, & initiates ITP.
Validated NA purity with off-chip qPCR.
Detection achieved over 2 orders of
magnitude parasite concentration.
Minimum detection = 500 parasites/µl
1st demo of on-chip integration of thermal
blood lysis and NA extraction.
1st demo of lysis and NA extraction with no
external actuation (no pump, no mixer, no
moving parts!)
Total nucleic acid (NA)
extracted from whole human
blood infected with P.
falciparum (malaria).
Compare to negative control to
verify parasite presence and
quantify concentration.
Standard glass capillary
PCB Lab-on-a-chip
Mark Bachman (UC Irvine) & Juan Santiago (Stanford)
Lab-on-a-Chip Commercialization
To date, process is long and cumbersome
– Many different impressive technologies, fabrication processes developed over the last 20 years.
– However, few successful commercial examples.
Trying to streamline this process….
– Consortium mechanism and partnering – work closely with industry – Application driven development – Foster the entrepreneurial spirit – provide resources, infrastructure – Use manufacturable processes that are already established,
characterized, etc.
– Ultimately develop design tools to expedite development
MANUFACTURING PROCESSES
USERS =
Universities,
companies,
govt. labs,
any designer
1) User designs device on computer via advanced software design tools
2) Design is then fabricated (on-site or via a network of off-site locations
depending on design). “Virtual foundries”
3) Utilize on-line community to test market and find collaborators for
packaging, marketing, pricing, etc.
Graphic Source: The Economist, April 21, 2012
SOFTWARE, DESIGN TOOLS
Future: “Digital” LOC Manufacturing
Conclusion: E-Health of the Future
Satellite Integrated, low-cost, simple
labs-on-a-chip that can
quickly perform assessment
anywhere and everywhere.
Ubiquitous Diagnostics
New NSF I/UCRC in 2014 = CADMIM
www.inrf.uci.edu/cadmim
Center for Advanced Design and
Manufacturing of Integrated Microfluidics