INTRODUCTION

MICROFLUIDIC DEVICE
KULIAH UMUM : SITI JULAIHA
HOW SMALL IS SMALL
PHYSICAL ASPECTS
 DEFINITION
Microstructure products :
-Structures in the micrometer range
-Have technical function provided by the shape of the
microstructure.

Microsystem combine several microcomponents, optimized as

an entire system, to provide one or several specific functions,
in many cases including microelectronics

NEXUS (The Network of Excellence in Multifunctional Microsystems) 1998

Europaeischer Industrieverband fuer MST
 MICROSYSTEM TECHNOLOGY
Signal generation and processing
Functional Materials : Transform information into signals or signal
into actuation
Handling of system components
Construction Materials :Mechanical stability, combination of
elements
Miniaturization
Use small amounts of e.g. liquid or energy

Integration
Various system components integrated in one system in close vicinity
from One base (monolithic integration) without or with little
mounting effort
Improved reliability, reduced cost and size

Microtechnological Manufacturing
Fabrication of many systems in parallel processes (batch fabrication)
 MICROSYSTEM TECHNOLOGY
• Apply methods of microelectronics to
• Micromechanics (sensors, gyros, drives, actuators, etc.)
• Microoptics (integrated optics, optical switches )
communication technologies)
• Microfluidic (gases , liquids) such as bio chips, Lab on a
chips, Inkjet printers, medical dosing, etc..)
• Important :
-Combining technologies
-Integration of functions
-Integrated manufacturing process
• Add classical mechanical technologies
Micromachining, micromilling, injection molding,
Hot embossing, laser structuring
 ADVANTAGES OF MICROSYSTEMS
• Less Connections
Improved reliabillity
• Miniaturization
New application areas,

Less mechanical load

• Redundancies possible
Improved reliability

• Integrated signal handling an processing

Compansation for error possible

Advanced functionality
• Reduced costs
Batch processing
• Autonomous systems
Low energy consumptions
• Fast Reaction times
Short distances

IN GENERAL : SMALL VOLUMES, LOW ENERGY CONSUMPTION

 WHAT IS FLUID?
A FLUID IS A MATERIAL (GAS OR LIQUID) THAT
DEFORMS CONTINUALLY UNDER SHEAR SHEET, i.e.,
THE MATERIAL CAN FLOW AND HAS NO RIGID 3-D
STRUCTURE
 MICROFLUID
FLUID IN EVERYDAY LIFE

HOW THE FLUID

BEHAVE IN A
REDUCED SCALE
MICROFLUID
SCALLING EFFECT IN MICROFLUID
• Laminar Flow (Low Reynold
Number)

• Electric Double Layer

• Surface forces become

dominant

• Reduction of Sample volume

• Liquid evaporation

•  Impurities and Bubbles

• Assembly Packaging and

Integration
IMPORTANT PARAMETER/PROPERTIES
DENSITY
 PRINCIPLES
PASCAL
Tekanan yang diberikan zat cair
dalam ruang tertutup diteruskan ke
segala arah dengan sama besar

ARCHIMEDES
Jika suatu benda dicelupkan ke
dalam sesuatu zat cair, maka
benda itu akan mendapat tekanan
keatas yang sama besarnya
dengan beratnya zat cair yang
terdesak oleh benda tersebut

Fa=ρ.g.V
VISCOSITY
Temperature Dependence of Viscosity
TURBULENCE AND REYNOLDS NUMBER
EXAMPLE Re IN A MICROSTRUCTURE
 A CLOSER LOOK LAMINAR FLOW vs Re #

• Transitional Reynold numbers of ca. 2300 were

found empirically for macrosocpic systems with
fully developed flow

• In parts of microsystems (e.g, in valves) the

available flow path might be shorter than the entry
length necessary to develop full flow

• Transitional Reynolds numbers might be much

lower than 2000 depending on geometrical factors

• Close examination and modelling/simulation

necessary
WHAT KIND OF FLOW GET IN MICROCHANNELS?

Microchannels
Re < 100 (often less than 1)

Flow is completely laminar and no turbulence occurs.
Laminar flow provides a means by which molecules
can be transported in a relatively predictable manner
through microchannels.
 Materials and Fabrication
Polymeric Laminate Technology
Materials: SILICON
BEST FOR CHANNELS COUPLE WITH
MICROELECTRONICS OR
MICROELECTROMECHANICAL SYSTEMS (MEMS)
Good stiffness,
Allowing formation rigid microstructures (dimensional stability).

Figure 1. Photographic image of Si-Pyrex microdevice (H-filter)

used in the Yager laboratory. It consists of a Si piece with
anisotropically etched channels and four mechanically drilled
through-holes, covered by a Pyrex cover that has been
anodically bonded for a hermetic seal.
Materials and Fabrication
 Materials and Fabrication
Polymeric Laminate Technology
Materials: POLYDIMETHYLSILOXANE (PDMS)
OPTIC CLEAR FLEXIBLE
(EASY STACK ONTO OTHER
CURED POLYMER SLABS)

FORMING COMPLEX 3D
GEOMETRIC
 Materials and Fabrication
Polymeric Laminate Technology
Materials: OTHERS

Material: Fabrication Technique:

Silicon Chemical wet etch
Glass Chemical etch, laser cutting

Polymeric films (e.g., Mylar) Laminate laser cutting

Silicone elastomer (PDMS) Micromolding ("soft lithography")

Photopolymerization ("microfluidic
Photoresist, hydrogels, etc.
tectonics")
Thermoplastic Hot embossing, injection molding
METHODOLOGY
 TWO COMMO# METHODS :
by which fluid actuation through microchannels can be achieved.

PRESSURE DRIVE# FLOW

Fluid pumped through the device via positive displacement pumps
(syringe pumps)
Basic Law :
The so-called no-slip boundary condition, states that the
fluid velocity at the walls must be zero.
This produces a parabolic velocity profile.

Figure 1.1. Velocity profile in a microchannel with aspect ratio 2:5 under
conditions of pressure driven flow. (ote that the velocity is assumed to be
zero at the walls in most treatments of transport of liquids. Image from
result of a calculation using Coventorware software.
 Electrokinetic Flow
Electroosmotic pumping.
Microchannel wall with an electric charge, forming an electric double
layer of counter ions .

Electric field applied :

- Across channel, ions in the double layer move towards the electrode of
opposite polarity. Creates motion fluid near the walls and transfers via
viscous forces into convective motion of the bulk fluid.
-Open channel at the electrodes, the velocity profile is uniform across the
entire width of channel.

- Closed channel, a recirculation pattern forms along center of the

channel moves in a direction opposite to that at the walls . The velocity
along centerline of channel is 50% of the velocity at the walls.
PDMS MicroFluid Device Process
Prepare PDMS
Mix 10:1 silicone
elastomer and curing
agent

Eliminate bubbles by
putting in vacuum
pump

Pour mixture in the

mold. Heat at 120oC.
28
PDMS MicroFluid Device Process
Laminating step
Clean glass slides and dry
properly

Laminate the photoresist

onto the glass WITHOUT
BUBBLES

Cover with the foil containing

the patterns

29
PDMS MicroFluid Device Process
Developing Process
Expose the pattern to
UV light.

Place in Na2CO3
solution to remove
unexposed area

Re-expose and heat the

pattern to harden it.
30
PDMS MicroFluid Device Process
Creating fluid supply
Add “needles” to the part
where liquid will be placed

Pour PDMS in the

mold with the pattern

Heat to harden
PDMS

31
PDMS MicroFluid Device Process
Making micochannel
Remove PDMS from
the glass pattern

Expose in oxygen
plasma for activation

Close the channels with

PDMS or glass lid.
32
PDMS MicroFluid Device Process
Testing and Analysis

Test microfluidic devices

made

Compare PDMS-PDMS
and PDMS-glass & the
two patterns we have

Observe the flow

dynamics through a
microscope
34
Two different liquids in the microchannel did
not mix.
Experimental Parameters

 Comparing 2 different systems: PDMS-

PDMS and PDMS-glass microfluidic
devices
 Comparison of velocity and flow rate with
and without syringe
 Flow dynamics (layering vs mixing of
liquids) in different systems
Measured the velocity of the fluid in the
microchannel

Fluid velocity (with syringe)

1.2 Possible Errors:
1
-Pushing the syringe
-Pressure in the tube
Velocity

0.8
-Water/bubbles in the
0.6 channel
0.4

0.2

0
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5

Time (s)
To minimize the errors, removed the syringe
and observed the flow

Fluid velocity without syringe

1.2

1
Velocity

0.8

0.6

0.4

0.2

0
0 0.5 1 1.5 2 2.5 3 3.5

Time
Some insights on the experiment

 The colored-liquid takes time to flow

because PDMS is hydrophobic as shown
by water-drop experiment.

Water

PDMS Glass
Insights during the process

 At the micro-level, Reynold’s number

is so low that fluids behave very
viscously and flow laminarly and they
do not mix.


 For the fluid, the highest Reynold’s

number (fastest velocity) is 2.36
Increasing length of channel allows for more
diffusion and mixing
Summary

 Microfluidic device is a new, convenient

and efficient tool for a lot of biological
applications.
 The device takes advantage of properties
in the micro- level such as the laminar flow
of fluids given by low Reynold’s numbers.
 Other advantages include small volume
size, easy-to-use, and wide variety of
applications.
Summary

 Fabrication of microfluidic device used the

photoresist and a mask as a template; and
PDMS was used as the molding agent.
 PDMS was used because the surface can
be modified by oxygen plasma so it can
form a Si – O – Si covalent binding with a
PDMS or Glass (SiO2) lid.
 Cleanliness, however, was critical
throughout the experiment.
 Conclusion

 Results show that fluids indeed behave in

a laminar manner if the Re number is low;
but longer channels allow time for diffusion
causing them to mix.
 Velocity decreases through time
quadratically in the pressure-driven flow
(syringe) and the capillary force-driven
flow.
 METHODS
Pattern Transfer to SiO2 thin film :
 METHODS
PHOTOLITOGRAPHY

Slow, poor adhesion,

Fast, good adhesion,
high resolution
swelling
 METHODS
ANISTOTROPIC ETCHING
 METHODS
ANISTOTROPIC ETCHING
 METHODS
SURFACE MICROMACHINING
MICROMILLING TOOS
MICROMILLING TOOS
MICROMILLING TOOS
MICROMILLING TOOS
LASER ABLATION
LASER ABLATION
LASER ABLATION
PATERNING OF CONDUTING POLYMERS WITH CO2 LASER
LASER WELDING
LASER WELDING
LASER WELDING
HOT EMBOSSING
HOT EMBOSSING
BONDING (POLYMERS)
OTHER APPLICATIONS
APPLICATIONS IN MEDICAL TECHNOLOGIES
Table I. Breakdown of Immunodiagnostics by Category
Application Share Analyte type Concentratio
n range
Drug testing 26% Small molecules 10 nM - mM
Therapeutic 20%
Drugs of abuse 6%
Infectious diseases 21% Bacteria, viruses, large low
STDs 13% molecules, antibodies and
Non STDs 8% metabolites
Endocrinology/hormones 33% Small molecules, large pM - nM
Thyroid 20% molecules including Abs
Non-thyroid 13% (70% <5 kDa)
Immunology 7% Antibodies pM - nM
Allergy 3%
Autoimmunity 3%
Other 1%
Cancer 5% Large molecules, pM - nM
antibodies
Other 8% > nM
Electrophoresis
This Movie show how Microfluidic Device could be used with
Electrophoresis with Direct Current for separation cancer cells

..\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic

Engineering\Electrophoresis\DC-Dielectrophoretic separation -
cancer cells [www.keepvid.com].mp4

And this to separate between sperm cells and the cells from skin
..\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic
Engineering\Electrophoresis\DC-Dielectrophoretic separation -
skin and sperm cells.mp4

And for separate cell in different diameter or shape

..\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic

Engineering\Electrophoresis\DC-Dielectrophoretic separation of
particles.mp4
 One of research in Biomedical application
Is from
Valero et al. “ Gene transfer and protein dynamics in stem cells using
Single cell eletroporation in a microfluidic device
..\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic
Engineering\Electrophoresis\Design, fabrication and cell-handling of
microfluidic device for single cell electroporation [.mp4

Sort particles based on their size and Electrical properties

..\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic
Engineering\Electrophoresis\Dielectrophoresis Explained
[www.keepvid.com].mp4
he sorted drops move up the field gradient created by the electrodes by
dielectrophoresis and are pulled into the keep channel. The movie is
recorded at 4,000 frames s-1 and shows the result of sorting a
demonstration emulsion containing fluorescein (light drops) or
bromophenol blue 1 percent by weight in water.
sorting a demonstration emulsion containing fluorescein (light drops) or
bromophenol blue 1.flv
 BIOMEDICAL APPLICATIO#

Diffusion Immunoassay (DIA)

Monitor Drug levels in body fluids,

Diagnose infectious and autoimmune diseases
Diagnose and monitor treatment of cancer.
Modeling suggests to detect molecules below 1-10 nM
range
Monitor concentrations of analytes as large as proteins
 Immunoassay
Imunoasai adalah tes biokimia yang mengukur konsentrasi bahan pada cairan
biologi,seperti plasma darah atau urin, menggunakan reaksi antibodi terhadap
antigen. Asai mengambil kelebihan melekatnya antibody terhadap antigen. Sntibody
monoklonal sering digunakan dan mereka hanya melekat pada satu situs molekul, dan
selanjutnya menyediakan tes yang lebih spesifik dan akurat, yang tidak
membingungkan oleh kehadiran molekul lainnya. Antibody yang diambil harus
memiliki afinitas terhadap antigen.

Example: Sandwich assay

Detection by antibody-
antibody-antigen
antigen--antibody
antibody--binding

inlet outlet

active surface
 Immunoassay

 prepare active surface with antibodies (catching molecules)

inlet outlet

active surface
 Immunoassay

 inject probe (incubation)

inlet outlet

active surface
 Immunoassay

 Wash

inlet outlet

active surface
 Immunoassay

 marked antibody (conjugated antibody) + incubation

(antigen acts as a „glue“
between marker and wall)

inlet outlet

active surface
 Immunoassay

 Wash  Detection by e.g.:

 Fluorescence
 Chemoluminescence
 various other markers (e.g. radioactive marker [RIA])

inlet outlet

active surface
 ..\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic
Engineering\Microarray\Microarrays [www.keepvid.com].mp4
Target

 manually or with pipetting robot

 complicated handling
 large and expensive „Hardware“
 not mobile

 development of cheap (disposable-) Chips

für biochemical analyses
 all complex components integrated in a non disposible cheap reader
device
Components

 for performing the reaction

 Reaktion chamber

 Transporting the reagents

 Pump mechanism

 combining the reagents

 channel crossing

 storing the reagents

 liquid reservoirs

 Prozess control
 Computer

 analysis of the reaction

 read out system
Mikrofluidic reaktion chambers

 manufacture reaction chambers

 insert immuno chemistry in the reaction chamber
 (directly or on a substrate)
 close structures with adhesive foil
 advantage: no temperature / solvent necessary
  no destruction of bio chemical reagents

 chemicals applied from outside by tubes

 SU8 Photoresist

belichten

Entwickler

Deckel
SU8 Struktur Maske
SU8 Basis
Opferschicht
Substrat

☺
Immuno chemistry

 Manufacture reaction chambers

 fill in immono chemistry
 close chambers by adhesive foli
 advantage: no temperature / solvant necessary
  no destruction of bio chemistry

Deckel Antikörper
Antikörper
auf Substrat
auf Substrat
Kanalstrukturen

☺
Adhesive foils

 like expected:
different foilsdifferent chemistry

Vergleich verschiedener Folien

Blanc Positive
1000000
RLU's * Buffer labeled CRP-Z labeled
Foil 1 41 4859 100000

Relative Light Units [arb.]

Foil 2 37 73289 10000
Foil 3 41 53893
Foil 4 37 295090 1000
Foil 5 27 224809 100
Foil 6 40 344121
10
Foil 7 35 455444
Foil 8 38 313930 1
* RLU = Relative Light Units 1 2 3 4 5 6 7 8
Foliennummer
Conclusion
Assay works

☺
Liquid transport

 active controllable system

 good control of liquid transport
  Pump
 disposable / integrable
 simple / robust
 no moving parts
 integrated in manufacturing process of the channels
No Moving Parts Valves (NMP Valves)

Membrane pump with fixed valve structures

 No-Moving-Part-Valves (NMP-Valves)
(different flow resistence by reversal of flow direction)
No Moving Parts Valves (NMP Valves)

Membrane pump with fixed valve structures

 No-Moving-Part-Valves (NMP-Valves)
(different flow resistence by reversal of flow direction)
Valve variants

round bristles straight bristles

S perr-Durchlass-Verhältnis der Dioden bei Druckänderung

1,4
1,3
1,2
1,1
1,0
0,9
0,8
gerade Borsten (150µm)
0,7
runde Borsten (150µm)
0,6
Kehrwert der runden Borsten
0,5
0 20 40 60 80 100 120 140
∆p [kPa]
 Simulation

check direction flow direction

(with the bristles) (against the bristles)

 Simulation

check direction flow direction

(with the bristles) (against the bristles)

Test setup of the pump

 setup of the pump

test setup of the pump

☺
Test setup of the pump

 pumping rates over 500 µl/min

Pumpaufbau mit SU-8 Dioden 400 µm Höhe

700
600
Pumprate [µl/min]

500
400
300
200
100 Pumprate SU-8 400µm
0
0 10 20 30 40 50
Anregungsfrequenz [Hz]

counterpressure up to 4 kPa
Integrable!

 Integration in einen Labchip

☺
Computercontrol with LabView®

☺
Liquid reservoirs

integrated in the chip

should be completely usable
should not allow bubbles
should hold liquid even when dropped
 long, „wound up“ channels
 hold liquid by capillary force

☺
Optimization

 Problem
 slow flow and bad liquid exchange
at the sides
Optimization

 Test:
 flow optimized chamber
Optimization

 Solution
 Nitrocellulose disc
  reduced chamber higth
at biologically active region
 longer reaction chamber
Optimized channel crossing

 Solution
 „Venturi nozzle“
 at low inlet pressure no
flow at the side channels
 at higher inlet pressure
suction on the side
channels
 Prototypchip

waste reaction
reservoir chamber

washing tracer
pump pump

washing- tracer
reservoir reservoir

crossing probe
injection
 Basics

Detection of a cardiac infarct

 Cardiacmarker
 Myoglobin
 normal concentration in blood < 75 ng/ml
 raised e.g. by cardiac infarct ~ 700 ng/ml

 also raised by other effects

backup by other markers

 Troponin
 Creatinkinease (CK-MB)
 C-reactive-protein (CRP)
Immunoassay for detection of cardiac infarct

 Fluorescence detection
 no additional components necessary
 can be performed directly in the chip
Myoglobin-Immunoassay in PMMA-Chips

Chip 1 Chip 2
9,5
Peakmaximum des Fluoreszenzsignals

erste Messreihe
9
erste Messreihe
8,5 zw eite
8 Messreihe zw eite
[arb. units]

Messreihe
7,5
7
6,5
6
5,5
5
0 100 1000 10 500 0 100 1000 10 500
Myoglobinkonzentration [ng/ml]
Idea  $$$


Idea  $$$



 
 
 
 ?
Next steps

Proof of principle achieved

 still a long way to a product and to the marked

 reproducibility
 calibration on chip
 detect several substances (multi analyte assay)
 mass production
 storage
 stability over time
 System development (reader device, control electronics, controlling software@)
 Risiko analysis (e.g. wrong positive / negative results)
 approvals (FDA,@)
 Marketing channels
 Introduction into marked
 Sales- and manufacturing partners
 @

 Microfluidics still has not brought that much products to the market
 Prototype  Series

 Mass Production of piezo-micro positioning drives is a solved

problem and can serve as example

 Positioning systems, drive-, product- and system development

 Prototype  low volume series  high volume production
(every step has individual challenges)
 Mass production concepts for micro product in high volumes demand for
specialized quality measures and controlling mechanisms

Produktion capacity
~1Mio Motors / Year

Production facility of
Working principle

 diriving by resonant micro steps

 friction based driving principle
 FWD @ 80kHz
 BWD @ 100kHz
 fast: 300mm/s !

 Precise: ~µm
 higher precision (subµ) feasible
 dynamic: start/stop < 5ms
 Low cost mass product

>100
Positions per
second!
Example video Elliptecmotor

X15G_Motorbewegung.WMV
Example video Elliptecmotor

Delta-20090417_detail.wmv
Example video Elliptecmotor
 Prototyp  Serial product

 next development steps based on experience from mass production

of piezo micro positioning drives
Research & development:
 typically customer driven or driven by new ideas
 Development for function and manufacturability (close customer- / user relation)
 System development  Interdisziplinary (Construction, Prototyping, Elektronics, Software, Produktion X)

Production:
 evaluation of all available data
(during produciton an in final tests)
 storage of all data in a database
 evaluation of the data for statistical production control
 further product development by correlation of data
 controlling production by production data
  „self teaching production“

Marketing:
 support customer with product application
 customer specific product development / refinement
 analysis of target markets and marketing channels
 Innovative marketing strategies for new products necessary
Thank you for your attention
 REFERENSI
• Prof. Dr.-Ing. Michael Schlüter , Development of a Lab on Chip Device and
Mass Production of Microstructure Products
• Prof Oliver Geschke, Technical University of Denmark, Microsystem
Engineering of Lab on a Chip Devices, Winheim , Germany
• Siti Julaiha Presentation, Summer School Biomedic, Wilhemshaven 2010
• A rapid diffusion immunoassay in a T-Sensor, Hatch, A., Kamholz, A.E.,
Hawkins, K.R., Munson, M.S., Schilling, E.A., Weigl, B.H. and Yager P., Nature
Biotechnology, 19(5), 461- 465 (2001)
• Microfluidic Device Mimics Tumor Microenvironment, Helps Drug
Discovery Effort,February 23, 2009 , Neil Forbes, Ph.D., and his
colleagues at the University of Massachusetts
• Microfluidics, Catherine Cabrera, Katerina Macounova, Ph.D., Mark Holl,
Ph.D., Eric Schilling, PY
• Applying microfluidic chemical analytical systems to imperfect samples, Yager,
P., Bell, D., Brody, J.P., Qin, D., Cabrera, C., Kamholz, A., and Weigl, B.H.
• In Micro Total Analysis Systems '98, D. J. Harrison and A. van den Berg, eds.,
Kluwer Academic Publishers, Dordrecht, 207-212 (1998)
• Design of microfluidic sample preconditioning systems for detection of
biological agents in environmental samples, Yager, P., Afromowitz, M.A., Bell,
D., Forster, F.K., Brody, J.P., Qin, D., Cabrera, C., Holl, M., Kamholz, A., and
Weigl, B.H., SPIE Proceedings, 3515, 252-259 (1998)