Microfluidics in life sciences Applications (I) · Lab on a chip concept: ... Large surface/volume...
Transcript of Microfluidics in life sciences Applications (I) · Lab on a chip concept: ... Large surface/volume...
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Microfluidics in life sciencesApplications (I)Joel S. Rossier
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2Rossier EPFL 2011
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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3Rossier EPFL 2011
What is micro? What is nano?
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4Rossier EPFL 2011
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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5Rossier EPFL 2011
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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6Rossier EPFL 2011
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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7Rossier EPFL 2011
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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8Rossier EPFL 2011
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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9Rossier EPFL 2011
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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10Rossier EPFL 2011
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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11Rossier EPFL 2011
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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12Rossier EPFL 2011
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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13Rossier EPFL 2011
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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14Rossier EPFL 2011
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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15Rossier EPFL 2011
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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16Rossier EPFL 2011
Surface based microfluidics
Electrowetting
For example Advanced Liquid
Logics
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17Rossier EPFL 2011
Electrowetting
Surface tension modulation by application of an electric field
http://www.liquavista.com
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18Rossier EPFL 2011
Electrowetting theory
http://en.wikipedia.org/wiki/Electrowetting
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19Rossier EPFL 2011
Fabrication of electrowettingsupport hardware
Printed circuit boardused to pattern electrodes underdielectrics
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20Rossier EPFL 2011
Electrowetting
Dielectric
Electrode
+++ - - -
Voltage
Hydrophobic coating
Electrostatic charges: hydrophilic
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21Rossier EPFL 2011
Electrowetting
Dielectric
Electrode
+++- - -
Voltage
Hydrophobic coating
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22Rossier EPFL 2011
Electrowetting
Dielectric
Electrode
- - - Hydrophobic coating
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23Rossier EPFL 2011
Electrowetting
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24Rossier EPFL 2011
Electrowetting: different designs are possible
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25Rossier EPFL 2011
Beads or cells can be transported
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26Rossier EPFL 2011
Advance Liquid Logics
ImmunoassayPCRDilution
http://www.liquid-logic.com/applications.html
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27Rossier EPFL 2011
Surface Accoustic wave as a pumpingmeans
Pressure transfered to the liquid
www.advalytics.com
Tsunami effectRayleigh Wave
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28Rossier EPFL 2011
Advalytics: surface accousticwave
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29Rossier EPFL 2011
Nanodrop : optical cell of 1 microLOptical cell held by surface tension1 drop of solution (1 microL)Spectroscopy in a drop
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30Rossier EPFL 2011
Nanodrop
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31Rossier EPFL 2011
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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32Rossier EPFL 2011
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33Rossier EPFL 2011
Pressure control with Fluigentsystem
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34Rossier EPFL 2011
Comparison of Syringe drivenflow and Fluigent system
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35Rossier EPFL 2011
Flow control by Fluigent
Oil drop formation Block flow Injection
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36Rossier EPFL 2011
Microfluidic separation
Capillary Electrophoresis Chip Electrophoresis MS sampling (nanoelectrospray) HPLC MS
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37Rossier EPFL 2011
Capillary electrophoresis
Separation method occuring in a capillary
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38Rossier EPFL 2011
Electro-induced convection
Electroosmotic flow Characteristics:
almost flat front in microchannels
In nanochannel: again a sort of parabolic profile!
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39Rossier EPFL 2011
Migration+electroosmotic flow separation
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40Rossier EPFL 2011
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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41Rossier EPFL 2011
How to inject a small plug?
Injection pattern
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42Rossier EPFL 2011
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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43Rossier EPFL 2011
1D electrophoresis
Biopolymer Sizing Sieving through a
polymer matrix 1D-gel
electrophoresis to be replaced by Chip electrophoresis
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44Rossier EPFL 2011
Caliper-Agilent technology Lab on a chip
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45Rossier EPFL 2011
Separation with Bioanalyser
View as a Gel or Electropherogram
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46Rossier EPFL 2011
Fully Automatic LOC
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47Rossier EPFL 2011
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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48Rossier EPFL 2011
ScreeTape, Lab 901:
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49Rossier EPFL 2011
Fluidigm: valving on a chip: system fully integrated
Application: qPCR
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50Rossier EPFL 2011
Fluidigm: valving with elastomer
Nanoflex Valve
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51Rossier EPFL 2011
How to inject into a microfluidicchip network
Pressure pinched injection
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52Rossier EPFL 2011
Plug with parabolic shape
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53Rossier EPFL 2011
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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54Rossier EPFL 2011
3-D organisation Optimisation of surface by nanopillars Microfabricated organised surface
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55Rossier EPFL 2011
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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56Rossier EPFL 2011
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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57Rossier EPFL 2011
Monolithic columnMonlithic column can befabricating by polymerisation of monomerby UV activation.
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58Rossier EPFL 2011
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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59Rossier EPFL 2011
CEC with monolithic polymerised column
Throckmorton, D. J., Shepodd, T. J., Singh, A. K., Anal.Chem. 2002, 74, 784–789.
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60Rossier EPFL 2011
HPLC (High performance liquid chromatography Standard system:
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61Rossier EPFL 2011
Agilent HPLC on a chip
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62Rossier EPFL 2011
Nanostream: parallel HPLC on each cartridges
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63Rossier EPFL 2011
Use of nano(micro-) particles
Location of the beadsRandom organisation3-D organisation
Magnetic vs non magnetic beadsDifferent surface materialsDifferent compositions
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64Rossier EPFL 2011
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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65Rossier EPFL 2011
Magnetic particles can containfluoroscent properties
http://www.trendbio.com.au/NFPstructure2.gif
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66Rossier EPFL 2011
Magnetic Nanoparticles
Monodisperse(Ademtech)
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67Rossier EPFL 2011
Magnetic nanoparticle arrays
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68Rossier EPFL 2011
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69Rossier EPFL 2011
3-D organisation of bead columnwith magnetic field
Magnetic field organised column-Renewable chromatography phase-DNA separation-Enzymatic digestion
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70Rossier EPFL 2011
DNA Separation by sieving Long DNA strains separation
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71Rossier EPFL 2011
Application of microfluidics for affinity assay Immunoassay ELISA (enzyme linked immunosorbent
assay DNA Hybridisation etc
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72Rossier EPFL 2011
Diffusion Immunoassay:
Dilution by diffusion
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73Rossier EPFL 2011
Diffusion based assays
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74Rossier EPFL 2011
H-filter
Paul Yager’s Group
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75Rossier EPFL 2011
Affinity assay: Immunoassay
Standard Immunoassay technologyELISA (Enzyme Linked
ImmunoSorbent Assay)ELFIA (Enzyme Linked
Fluorescent ImmunoAssay)
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76Rossier EPFL 2011
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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77Rossier EPFL 2011
Diffusion Immunoassay II
YY Y Y
Y Y Y Y
Slow diffusing antibodyRapid diffusing antigen
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78Rossier EPFL 2011
YY Y Y
Y Y Y Y
Diffusion Immunoassay III
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79Rossier EPFL 2011
YY
Y Y
YY
Y Y
Diffusion Immunoassay IV
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80Rossier EPFL 2011
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.