microfluidics in life sciences applications (iii)€¦ · 2 rossier epfl 2011 multiplexing...
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Microfluidics in life sciencesApplications (III)Joel S. Rossier
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2Rossier EPFL 2011
Multiplexing
Microarray of DNAAffimetrix; Exonhit (full genome on a chip)
Microarray for proteinZeptosens, Zyomix (difficult to get a clear
vision of the market)
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Zeptosens
Planar waveguide detection Detection of fluorescent molecule attached
to the surface
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Zeptosens
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Zeptosens application
Reverse array: Immobilisation of
separatedcomplexedprotein mixture
Western blotting Identification
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Merger of microfluidics andmicroarray: a foot in the future
Example of paraflow
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Merger of microfluidics andmicroarray: a foot in the future
Example of paraflow
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SpinX technologies
Programmable fluidics
Thousands of assay in a cartridge
Valving by laser opening
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Introducing the SpinX gStack
Dose response curvesfor full assay panel:multiple compoundsxmultiple assaysxmultiple concentrations
Dose response curvesfor full assay panel:multiple compoundsxmultiple assaysxmultiple concentrations
Same system for:• more cpds, less assays• more assays, less cpds• assay development• hit-to-IC50 follow-up• …
Same system for:• more cpds, less assays• more assays, less cpds• assay development• hit-to-IC50 follow-up• …
Compounds(up to 100% DMSO)
Enzymes(stock solution)
Substrates(stock solution)
Other: buffer, ATP,detection reagents,…
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Complex Assays on the Bench-Top
Add AssayComponents
Load gStack
Analyze Data
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Programmable MicrofluidicsKinase“Hits”
KinaseIC50
ProteaseIC50
Kinase
KinaseBuffer Compound
DMSO
Substrate
ATP
Pierce IQ
Incubate
Readout
Protease
AssayBuffer
ProteaseBuffer
Compound
Substrate
Pre-incubate
Kinetic Readout
DyeBuffer
DMSO
Dilution of fluorescent dye
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Virtual Laser Valvecentrifugal force
VLVlocation defines volume metered
Single source chamber serves many destination chambers
Addition timing independent of reagent properties or amount of centrifugal force
sample A
sample B
reagent X
reagent Y
substrate
A-X A-Y B-X B-Y
Substrates
Microfluidic structures
Thin film which can be perforated by laser
SIDE VIEWFluid
Laser
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The SpinX Lab
gCards automaticallyloaded from gStack…
… to rotor
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Fluorescent Readout for Kinetics / End-Point
ContinuousRotation
Optical Modules based on DVD Pickup• VLV module: near-infrared laser + focus feedback• Readout modules (FI / FRET / TR-FRET, wavelength)
Ex. Fluorescence Intensity• Laser-based excitation• Emission photon counting
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Protein Crystallisation in microchip (Emerald bioscience)
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Protein Crystallisation in microchip (Emerald bioscience)
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Microfluidics : large surface to volume ratio “God made the bulk; the surface was
invented by the devil.”
Wolfgang Pauli
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Microfluidics in the air?
Droplet reactor No need for microfabricated device Reaction in a drop
Levitated Droplet
Prof Staffan Nilson; Lund University
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How to work with a levitateddrop ? Accoustic wave
Acoustic levitation uses sound pressure to allow objects to float.
http://static.howstuffworks.com/gif/acoustic-levitation-2.jpg
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Airborne Cell Chemistry
Staffan Nilsson www.sgms.ch/meeting/2007/Abstracts/S_Nilsson.jpg
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Airborne Cell Chemistry
Levitated separation. Right photo shows how one of the phases is discarded from the drop with a micropipette.
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Conclusion
Different technologies have been developed in the last 20 years
Products are coming with more or less success on the market