Microfluidics in life sciencesApplications (I)Joel S. Rossier

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Table of content

Why: Motivation for miniaturisation How: Design and Fabrication Where: Example of academic systems What: Example of industrial applications

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What is micro? What is nano?

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1. Why: Motivation for minaturisation

1. Small is beautiful?2. Small is cheap?3. Small is fashionable?4. Small is flexible?5. Small is …?

1. Smart, rapid, modular, portable, etc2. Mimic the example of electronic evolution with

Moore’s law.

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Progress of a technology - Electronic chips

Jack Kilby (TI) Nobel PrizeRobert Noyce (Intel) $$$$!

Texas Instr IC

1948 1958 1968 1971 2000

Transistor INTEL

Pentium 4

INTEL 4004 MP

Slides provided byLarry KrickaChairman of AACC

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Moore’s Law

Miniaturisation = performance^2

Each 18-24 months the performance is doubling and the miniaturisation

Cost down Now : 40 nm chips

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Progress of a new technology –Microchips: microarrays (Affymetrix)

FDA approvalRoutine use

ProductsOther labs

1st publicationPatents filed

Number of publications

1995 2002~25 ~1000

1982 ~1990 ~2000 2004e.g., GLC (Terry)

Slides provided byLarry KrickaChairman of AACC

RocheFDA

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Progress of a new technology - Bioelectronic chips (outside of Glucose sensing)

FDAapproval /// just i-STAT ///Routine use

ProductsOther labs

1st publicationPatents filed

1988 1993 ~2000 2004i-STAT Nanogen AVIVAMolecular Devices

Slides provided byLarry KrickaChairman of AACC

DiagnoSwiss

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Lab on a chip concept:

Has existed as a concept for about 15 years

Takes time to take off (Academic effort) Is now oriented towards classical

applications Has the potential to be applied in many

fields

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Flux of molecules:

The flux Ji of species i is given by the Nernst Plank equation as follow:

Ji Di grad ci

ziFRT

Dici grad ci (II.1)

where Di is the diffusion coefficient, ci the concentration, zi the charge and gradci the

concentration gradient of the species i, respectively and where grad is the potential gradient

in the solution, and where F, R, T and are the Faraday constant, the gas constant, the

temperature and the hydrodynamic flow respectively.

Diffusion Migration Convection

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Fields of applications for microfluidics Standard technics

Capillary electrophoresis HPLC Dispensing Mass spec. Infusion Injection (mosquito skin) Biosensor ELISA Ion channel detection MicroSynthesis …

Method only possible thanks to miniaturisation Single cell readout/feeding Power flow cell Power bio cell Neuron connection

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Fundamentals of fluidic behaviour in microenvironment < 50 microns Large surface/volume ratio Surface treatment becomes fundamental Fluid viscosity is important ! Solid particlesCoagulation, suspension etc

Gas handling (attention material permeation)

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Fluid characteristics

In microchannels:Low Reynolds/Peclet numberReduced natural convectionElectrokinetics pumpingMixing only by diffusion

Mixing efficency depends on the molecules parameters (size, Stokes/Einstein friction module)

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Surface treatment of polymer

Example with PET

Contact angle measurement

http://psii.kist.re.kr/Teams/psii/research/Con_4.jpghttp://web.mit.edu/nnf/people/jbico/drop.jpg

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Super hydrophobic surface with or without 3-D surface Nature Materials 1, 14–15 (2002)

Behaviour of water drops on different surfaces. a, Water drop wetting a normal surface forms a low internal contact angle. b, Non-wetting, quasi-spherical drop forms a high internal contact angle on a superhydrophobic surface. c, Drop on a solid surface decorated with pillars. The entire surface is coated with a water-repellent agent, and the space between the pillars is filled with air. Yoshimitsu et al.2 observe that the drop can sit happily on top of these pillars — the so-called 'fakir regime' —corresponding to an apparent contact angle larger than 150°(superhydrophobic behaviour). If the pillar height is shortened, the water contact angle decreases, because air is no longer trapped below the drop.

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Surface based microfluidics

Electrowetting

For example Advanced Liquid

Logics

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Electrowetting

Surface tension modulation by application of an electric field

http://www.liquavista.com

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Electrowetting theory

http://en.wikipedia.org/wiki/Electrowetting

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Fabrication of electrowettingsupport hardware

Printed circuit boardused to pattern electrodes underdielectrics

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Electrowetting

Dielectric

Electrode

+++ - - -

Voltage

Hydrophobic coating

Electrostatic charges: hydrophilic

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Electrowetting

Dielectric

Electrode

+++- - -

Voltage

Hydrophobic coating

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Electrowetting

Dielectric

Electrode

- - - Hydrophobic coating

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Electrowetting

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Electrowetting: different designs are possible

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Beads or cells can be transported

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Advance Liquid Logics

ImmunoassayPCRDilution

http://www.liquid-logic.com/applications.html

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Surface Accoustic wave as a pumpingmeans

Pressure transfered to the liquid

www.advalytics.com

Tsunami effectRayleigh Wave

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Advalytics: surface accousticwave

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Nanodrop : optical cell of 1 microLOptical cell held by surface tension1 drop of solution (1 microL)Spectroscopy in a drop

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Nanodrop

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Pressure driven flow on a chip

Fluidics without Electical field Mechanical pressure in the chip Open channel or packed channel for

affinity or chromatography separation

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Pressure control with Fluigentsystem

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Comparison of Syringe drivenflow and Fluigent system

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Flow control by Fluigent

Oil drop formation Block flow Injection

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Microfluidic separation

Capillary Electrophoresis Chip Electrophoresis MS sampling (nanoelectrospray) HPLC MS

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Capillary electrophoresis

Separation method occuring in a capillary

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Electro-induced convection

Electroosmotic flow Characteristics:

almost flat front in microchannels

In nanochannel: again a sort of parabolic profile!

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Migration+electroosmotic flow separation

+

-

+

-

+

- +

-

+

- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -

+ + + +

-

+

+

+ + + +

-

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Microfluidics vs Microvalving

Remote valving Electrokinetic injection Pressure control

In situ valving Local material deformation Material shifts

Microvalving is a challenge in microfluidics

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How to inject a small plug?

Injection pattern

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Microchip Structures for Submillisecond Electrophoresis

Figure 1 Schematic of microchip used for high-speed electrophoreticseparations. (Inset) Enlargement of the injection valve and separation channel.

Copyright © 2007 American Chemical Society

Anal. Chem., 70 (16), 3476 -3480, 1998

Figure 3 High-speed electropherogram of rhodamine B and di-chlorofluorescein resolved in 0.8 ms using a separation field strength of 53 kV cm-1 and a separation length of 200 m. The start time is marked with an arrow at 0 ms

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1D electrophoresis

Biopolymer Sizing Sieving through a

polymer matrix 1D-gel

electrophoresis to be replaced by Chip electrophoresis

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Caliper-Agilent technology Lab on a chip

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Separation with Bioanalyser

View as a Gel or Electropherogram

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Fully Automatic LOC

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ScreenTape: rapid electrophoresis

Similar but not identical!!! to Caliper/Agilent technology:Older microfluidics patents are going to expire (first work in late 80’s!) leading toMore similar products.

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ScreeTape, Lab 901:

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Fluidigm: valving on a chip: system fully integrated

Application: qPCR

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Fluidigm: valving with elastomer

Nanoflex Valve

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How to inject into a microfluidicchip network

Pressure pinched injection

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Plug with parabolic shape

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Nanochromatography

Packing or monolithicPacking

Easy to choose the surface of interest Tricky to immobilised in the nanocolum Use of frit or size restriction in the column

Monolithic In situ synthesis Lower choice of surface material Easier to adapt to any form

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3-D organisation Optimisation of surface by nanopillars Microfabricated organised surface

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Monolithic pillar for chromatography or CEC

Slentz, B. E., Penner, N. A., Regnier, F. E., J. Chromatogr. A2002, 948, 225–233.

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Microfluidic enrichment for low concentration (pM)

5. Jemere AB, Oleschuk RD, OuchenF, Fajuyigbe F, Harrison DJ:An integrated solid-phase extraction system for sub-picomolardetection. Electrophoresis 2002, 23:3537-3544.

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Monolithic columnMonlithic column can befabricating by polymerisation of monomerby UV activation.

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http://www.orc.wur.nl/UK/Events/PhD+trip+2007/Abstracts/Kishore+Tetala/

Scheme 1. Procedure to immobilize carbohydrates onto monolith beds: a) Original polymer bed with DATD as cross linker; b) 1,2-Diol group of DAD was cleaved using sodium per-iodate & c) Sugar with amino spacer was immobilized via reductive amination.

a) b) a) b)Figure 1. Optical microscope images: Figure 2. Scanning electron microscope images:

a) Empty column b) Monolith capillary column a) Empty capillary b) Monolith capillary column

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CEC with monolithic polymerised column

Throckmorton, D. J., Shepodd, T. J., Singh, A. K., Anal.Chem. 2002, 74, 784–789.

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HPLC (High performance liquid chromatography Standard system:

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Agilent HPLC on a chip

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Nanostream: parallel HPLC on each cartridges

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Use of nano(micro-) particles

Location of the beadsRandom organisation3-D organisation

Magnetic vs non magnetic beadsDifferent surface materialsDifferent compositions

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Magnetic nanoparticles

Different features Size dispersion

Monodisperse

Magnetic core Different magnetic core

diameter/volume ratioMagnetic nanoparticles vs microparticles-Higher surface to volume ratio-lower sedimentation

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Magnetic particles can containfluoroscent properties

http://www.trendbio.com.au/NFPstructure2.gif

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Magnetic Nanoparticles

Monodisperse(Ademtech)

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Magnetic nanoparticle arrays

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3-D organisation of bead columnwith magnetic field

Magnetic field organised column-Renewable chromatography phase-DNA separation-Enzymatic digestion

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DNA Separation by sieving Long DNA strains separation

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Application of microfluidics for affinity assay Immunoassay ELISA (enzyme linked immunosorbent

assay DNA Hybridisation etc

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Diffusion Immunoassay:

Dilution by diffusion

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Diffusion based assays

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H-filter

Paul Yager’s Group

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Affinity assay: Immunoassay

Standard Immunoassay technologyELISA (Enzyme Linked

ImmunoSorbent Assay)ELFIA (Enzyme Linked

Fluorescent ImmunoAssay)

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Diffusion Immunoassay in microfluidics

Hatch, A., Kamholz, A. E., Hawkins, K. R., Munson, M. S.,Schilling, E. A., Weigl, B. H., Yager, P., Nat. Biotechnol.2001, 19, 461–465.

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Diffusion Immunoassay II

YY Y Y

Y Y Y Y

Slow diffusing antibodyRapid diffusing antigen

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YY Y Y

Y Y Y Y

Diffusion Immunoassay III

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YY

Y Y

YY

Y Y

Diffusion Immunoassay IV

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Diffusion Immunoassay

Hatch, A., Kamholz, A. E., Hawkins, K. R., Munson, M. S.,Schilling, E. A., Weigl, B. H., Yager, P., Nat. Biotechnol.2001, 19, 461–465.