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