INTRODUCTION MICROFLUIDIC DEVICE KULIAH UMUM : SITI JULAIHA

HOW SMALL IS SMALL

PHYSICAL ASPECTS

in many cases including microelectronics NEXUS (The Network of Excellence in Multifunctional Microsystems) 1998 Europaeischer Industrieverband fuer MST . to provide one or several specific functions. Microsystem combine several microcomponents. optimized as an entire system.DEFINITION Microstructure products : -Structures in the micrometer range -Have technical function provided by the shape of the microstructure.

combination of elements Miniaturization Use small amounts of e.g.MICROSYSTEM TECHNOLOGY Signal generation and processing Functional Materials : Transform information into signals or signal into actuation Handling of system components Construction Materials :Mechanical stability. reduced cost and size Microtechnological Manufacturing Fabrication of many systems in parallel processes (batch fabrication) . 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.

. actuators. optical switches ) communication technologies) • Microfluidic (gases . Hot embossing. gyros. drives. Inkjet printers. etc.) • Important : -Combining technologies -Integration of functions -Integrated manufacturing process • Add classical mechanical technologies Micromachining. medical dosing.) • Microoptics (integrated optics. injection molding. micromilling. etc. laser structuring .MICROSYSTEM TECHNOLOGY • Apply methods of microelectronics to • Micromechanics (sensors. Lab on a chips. liquids) such as bio chips.

Less mechanical load Improved reliability • Integrated signal handling an processing Compansation for error possible Advanced functionality • Reduced costs • Autonomous systems • Fast Reaction times Batch processing Low energy consumptions Short distances IN GENERAL : SMALL VOLUMES.ADVANTAGES OF MICROSYSTEMS • Less Connections • Miniaturization • Redundancies possible Improved reliabillity New application areas. LOW ENERGY CONSUMPTION .

e.. i.WHAT IS FLUID? A FLUID IS A MATERIAL (GAS OR LIQUID) THAT DEFORMS CONTINUALLY UNDER SHEAR SHEET. 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 .

g.V . maka benda itu akan mendapat tekanan keatas yang sama besarnya dengan beratnya zat cair yang terdesak oleh benda tersebut Fa=ρ.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.

VISCOSITY .

Temperature Dependence of Viscosity .

TURBULENCE AND REYNOLDS NUMBER .

EXAMPLE Re IN A MICROSTRUCTURE .

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 necessary and modelling/simulation .A CLOSER LOOK LAMINAR FLOW vs Re # • Transitional Reynold numbers of ca.g. 2300 were found empirically for macrosocpic systems with fully developed flow • In parts of microsystems (e.

Laminar flow provides a means by which molecules can be transported in a relatively predictable manner through microchannels.WHAT KIND OF FLOW GET IN MICROCHANNELS? Microchannels Re < 100 (often less than 1) Flow is completely laminar and no turbulence occurs. .

covered by a Pyrex cover that has been anodically bonded for a hermetic seal. Allowing formation rigid microstructures (dimensional stability).Materials and Fabrication Polymeric Laminate Technology Materials: SILICON BEST FOR CHANNELS COUPLE WITH MICROELECTRONICS OR MICROELECTROMECHANICAL SYSTEMS (MEMS) Good stiffness. 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.

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 .

injection molding .g. etc.Materials and Fabrication Polymeric Laminate Technology Materials: OTHERS Material: Silicon Glass Polymeric films (e.. laser cutting Laminate laser cutting Micromolding ("soft lithography") Photopolymerization ("microfluidic tectonics") Hot embossing. Thermoplastic Fabrication Technique: Chemical wet etch Chemical etch. Mylar) Silicone elastomer (PDMS) Photoresist. hydrogels.

METHODOLOGY .

ote that the velocity is assumed to be zero at the walls in most treatments of transport of liquids.1. states that the fluid velocity at the walls must be zero. Velocity profile in a microchannel with aspect ratio 2:5 under conditions of pressure driven flow. Image from result of a calculation using Coventorware software. .TWO COMMO METHODS : by which fluid actuation through microchannels can be achieved. Figure 1. PRESSURE DRIVE FLOW Basic Law : The so-called no-slip boundary condition. Fluid pumped through the device via positive displacement pumps (syringe pumps) This produces a parabolic velocity profile.

The velocity along centerline of channel is 50% of the velocity at the walls.Closed channel. . .Across channel. -Open channel at the electrodes. Microchannel wall with an electric charge. ions in the double layer move towards the electrode of opposite polarity. Electric field applied : . a recirculation pattern forms along center of the channel moves in a direction opposite to that at the walls . forming an electric double layer of counter ions .Electrokinetic Flow Electroosmotic pumping. Creates motion fluid near the walls and transfers via viscous forces into convective motion of the bulk fluid. the velocity profile is uniform across the entire width of channel.

28 .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.

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. 30 . Place in Na2CO3 solution to remove unexposed area Re-expose and heat the pattern to harden it.

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 .

32 .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.

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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 .

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.Two different liquids in the microchannel did not mix.

Experimental Parameters Comparing 2 different systems: PDMSPDMS 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 1.5 2 2.4 0.8 0.5 1 1.6 Fluid velocity (with syringe) Possible Errors: -Pushing the syringe -Pressure in the tube -Water/bubbles in the channel 0.2 0 0 0.5 Time (s) .5 3 3.2 1 Velocity 0.5 4 4.

5 2 2. removed the syringe and observed the flow Fluid velocity without syringe 1.8 0.5 3 3.2 0 0 0.5 1 1.6 0.2 1 Velocity 0.5 Time .To minimize the errors.4 0.

Water PDMS Glass .Some insights on the experiment The colored-liquid takes time to flow because PDMS is hydrophobic as shown by water-drop experiment.

36 . Reynold’s number is so low that fluids behave very viscously and flow laminarly and they do not mix.Insights during the process At the micro-level. the highest Reynold’s number (fastest velocity) is 2. For the fluid.

Increasing length of channel allows for more diffusion and mixing .

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

and PDMS was used as the molding agent. Cleanliness. 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.Summary Fabrication of microfluidic device used the photoresist and a mask as a template. . was critical throughout the experiment. however.

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.Conclusion Results show that fluids indeed behave in a laminar manner if the Re number is low.

METHODS Pattern Transfer to SiO2 thin film : .

METHODS
PHOTOLITOGRAPHY

Slow, poor adhesion, high resolution

Fast, good adhesion, 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 .

nM > nM . antibodies pM .nM pM .Table I.nM Large molecules. antibodies and 8% metabolites 33% Small molecules. viruses. large 13% molecules. Breakdown of Immunodiagnostics by Category Application Drug testing Therapeutic Drugs of abuse Infectious diseases STDs Non STDs Endocrinology/hormones Thyroid Non-thyroid Immunology Allergy Autoimmunity Other Cancer Other Share 26% 20% 6% Analyte type Small molecules Concentratio n range 10 nM . large 20% molecules including Abs 13% (70% <5 kDa) 7% 3% 3% 1% 5% 8% Antibodies low pM .mM 21% Bacteria.

..\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic Engineering\Electrophoresis\DC-Dielectrophoretic separation cancer cells [www.\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic Engineering\Electrophoresis\DC-Dielectrophoretic separation of particles.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 skin and sperm cells.com].mp4 And this to separate between sperm cells and the cells from skin .mp4 And for separate cell in different diameter or shape .keepvid.mp4 .

fabrication and cell-handling of microfluidic device for single cell electroporation [..\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic Engineering\Electrophoresis\Dielectrophoresis Explained [www.flv .keepvid. The movie is recorded at 4.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 . sorting a demonstration emulsion containing fluorescein (light drops) or bromophenol blue 1.mp4 he sorted drops move up the field gradient created by the electrodes by dielectrophoresis and are pulled into the keep channel..mp4 Sort particles based on their size and Electrical properties .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.\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic Engineering\Electrophoresis\Design.com].

Modeling suggests to detect molecules below 1-10 nM range Monitor concentrations of analytes as large as proteins . Diagnose infectious and autoimmune diseases Diagnose and monitor treatment of cancer.BIOMEDICAL APPLICATIO Diffusion Immunoassay (DIA) Monitor Drug levels in body fluids.

dan selanjutnya menyediakan tes yang lebih spesifik dan akurat. Sntibody monoklonal sering digunakan dan mereka hanya melekat pada satu situs molekul. yang tidak membingungkan oleh kehadiran molekul lainnya. Asai mengambil kelebihan melekatnya antibody terhadap antigen. Antibody yang diambil harus memiliki afinitas terhadap antigen.Immunoassay Imunoasai adalah tes biokimia yang mengukur konsentrasi bahan pada cairan biologi.seperti plasma darah atau urin. Example: Sandwich assay Detection by antibody-antigen-antibody-binding antibody-antigen-antibody- inlet outlet active surface . menggunakan reaksi antibodi terhadap antigen.

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 .

mp4 .Immunoassay Wash Detection by e.com].: Fluorescence Chemoluminescence various other markers (e.g..g.\2_Bioteknologi & Rekayasa Sel & Jaringan\Genetic Engineering\Microarray\Microarrays [www.keepvid. radioactive marker [RIA]) inlet outlet active surface .

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 SU8 Basis Maske 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 Relative Light Units [arb.] Blanc Positive RLU's * Buffer labeled CRP-Z labeled Foil 1 41 4859 Foil 2 37 73289 Foil 3 41 53893 Foil 4 37 295090 Foil 5 27 224809 Foil 6 40 344121 Foil 7 35 455444 Foil 8 38 313930 * RLU = Relative Light Units Vergleich verschiedener Folien 1000000 100000 10000 1000 100 10 1 1 2 3 4 5 6 Foliennummer 7 8 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) .

5 0 20 40 60 ∆p [kPa] 80 100 120 140 gerade Borsten (150µm) runde Borsten (150µm) Kehrwert der runden Borsten .8 0.4 1.0 0.7 0.6 0.3 1.9 0.1 1.Valve variants round bristles straight bristles S perr-Durchlass-Verhältnis der Dioden bei Druckänderung 1.2 1.

Simulation check direction (with the bristles) flow direction (against the bristles) .

Simulation check direction (with the bristles) flow direction (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 l/min Pumpaufbau mit SU-8 Dioden 400 m Höhe 700 Pumprate [ l/min] 600 500 400 300 200 100 0 0 10 20 30 40 50 Pumprate SU-8 400 m 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 reaction chamber tracer pump waste reservoir washing pump washingreservoir crossing tracer reservoir probe injection .

by cardiac infarct ~ 700 ng/ml also raised by other effects backup by other markers Troponin Creatinkinease (CK-MB) C-reactive-protein (CRP) .g.Basics Detection of a cardiac infarct Cardiacmarker Myoglobin normal concentration in blood < 75 ng/ml raised e.

5 5 0 100 1000 0 100 1000 10 500 Myoglobinkonzentration [ng/ml] 10 500 Chip 1 erste Messreihe Chip 2 erste Messreihe zw eite Messreihe Peakmaximum des Fluoreszenzsignals [arb.5 8 7.5 9 8. units] zw eite Messreihe .5 6 5.5 7 6.Immunoassay for detection of cardiac infarct Fluorescence detection no additional components necessary can be performed directly in the chip Myoglobin-Immunoassay in PMMA-Chips 9.

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: ~ m higher precision (sub ) 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. Produktion 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 ) . Prototyping. Elektronics. Software./ user relation) System development Interdisziplinary (Construction.

Thank you for your attention .

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