microfluidics in life sciences applications (iii)€¦ · 2 rossier epfl 2011 multiplexing...

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