Introduction to Microfluidics4 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006 Why...

Introduction to Microfluidics

C. Fütterer, Institut Curie & Fluigent SA, Paris

Miniaturisation & Integration

3 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

MicroMicro--PipettesPipettes

Problems:• Minimal volume: 1μl • Samples unprotectedagainst evaporation & contamination (dust)• Slow and tedious operation• Manipulation errors• Difficult automationthrough expensive robots


Why TraditionalWhy Traditional Instruments are Instruments are UnsufficientUnsufficient

• Complexity: 100.000 genes and more than 106 proteins in human body.

•Spatio-temporal dynamics of cellular/molecular processes.

• Big volumes: low speed of thermal and molecular diffusion and reactions.


Size of Size of Biological ObjectsBiological Objects

nlpl mlμl

?


Solution: Solution: MicrofluidicsMicrofluidicsCome CloserCome Closer toto Cells and MoleculesCells and Molecules!!

Smallest fluid quantities (single cell)High throughput (statistics)Faster physico-chemical processes (diffusion more efficient)

Towards quantitative biology!

Institut Curie


Pioneer Pioneer PublicationPublication

Electronic Chips


Chips are not Chips...Chips are not Chips...

Institut Curie

DNA-Chips

Electronic Chips Microfluidic Chips


Many Advantages Many Advantages of Miniaturisation of Miniaturisation

• Size: nanoliters, single cell and molecule manipulations,• Superior flow control (no mixing),• Speed: diffusion: 1mm -> 15 min, 10μm -> 100ms),• Integration & parallelisation,• Protection against contamination and evaporation,• Kinetics easy to study.


- Arteria and venes in animals- Capillaries in plants

Microfluidics is Microfluidics is Not NewNot New

How to Make it:Microfabrication


PhotolithographyPhotolithography

•Spincoat photosensitive solution,•Dry•Put mask on top•Expose to UV•Wash•Heat• Master

PROTOCOL

Mask:


Rapid PrototypingRapid Prototyping: : PrinciplePrinciple

Chip-Fabrication

Master

MasterChip (PDMS)

Surface-Patterningby μ Contact Printing


Rapid PrototypingRapid Prototyping: : Mask and MouldMask and Mould

Chip-Fabrication

Chip (PDMS)


Rapid PrototypingRapid Prototyping: : MouldMould

Chip-Fabrication

Chip (PDMS)


MouldMould: : VialsVials

Reproducible and clean vials

Chip (PDMS)

Master

Air

Polymer

Microstructure


Rapid PrototypingRapid Prototyping: : Master Master II

Chip-Fabrication

Chip (PDMS)

Pallandre, Fütterer et al., 2006

Much more stable!


Rapid PrototypingRapid Prototyping: : Master Master IIII

Chip-Fabrication

Chip (PDMS)


Rapid PrototypingRapid Prototyping: Polyester : Polyester MasterMaster

Chip (PDMS)


TayloredTaylored Channel Channel SystemsSystems, , IntegrationIntegration

Quake et al.

Flow Control


MicroMicro--FluidicsFluidics: : Defined Flow Defined Flow ProfileProfile

Laminar profile


Difficulty: Flow Control through Interfacing Difficulty: Flow Control through Interfacing (Contraction)(Contraction)

Humanscale

Molecules andCells1cm

10μm

Ratio: 106 !!!


FlowFlow Control: Syringe Control: Syringe PumpsPumps

Syringe pumps:• Long response time • Strong hysteresis• Limited volume• Stopped-flow/flow inversion difficult

Elasticity

Bubbles Bending of tubesAtmospheric Pressure

SlowMechanical drive


FlowFlow Control: Control: Electroosmotic PumpsElectroosmotic Pumps

• Reproducibility problems with biological fluids

• Time consuming preparation

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

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

+ + + + + + + + + + + + + + +

+ + + + + + + + + + + + + + + + -


FlowFlow Control: ACControl: AC--Electroosmotic PumpsElectroosmotic Pumps

• Reproducibility problems with biological fluids

• Time consuming preparation• Electrodes are consumed muTAS 2005


FlowFlow Control: Control: Microfabricated PumpsMicrofabricated Pumps

• Pulsed flow• Complex Microfabrication• Objects are squeezed (Cells!)

Pressure

Quake et al.


FlowFlow ControlControl

• Peristaltic pumps

• Pulses• Long time constant• Objects are squeezed• Tubes have to be exchanged


Capillary Forces or Centrifugal SystemsCapillary Forces or Centrifugal Systems

Blood test Separation of blood plasma(Zengerle et al. IMTEK)


Pumping through Moving ObjectsPumping through Moving Objects

Leach et al. Lab Chip 2006

Opticaltweezer

Opticaltweezer


New New PrinciplePrinciple for Microfluidics for Microfluidics FlowFlow ControlControl

Microchannel

Calibrated leaks

“Semi-open” pneumatic pressure control

“PneumaticWheatstone bridge“

Microchannel

Calibrated leaks


New New PrinciplePrinciple for Microfluidics for Microfluidics FlowFlow ControlControl

“Semi-open” pneumatic pressure control

Very short response time (< 1s),

No pulsations,

Symmetry: flow in both directions,

Dynamic pressure feedback:automated compensation of

- back pressures,- bubbles, - environmental pressure

changes.


Vision of the Vision of the LabLab

today tomorrow

Microfluidic Chip:


MMicroicroFFluidicluidic CControl ontrol SSystem ystem StartStart--up up ““FluigentFluigent””

Industrial prototype

Pressure output

20cm

www.fluigent.com


Very Fast Response in Very Fast Response in MicrochannelMicrochannel

single T4 DNAmolecule

playing with sliding knob

Applications


Low Dispersion SuspensionsLow Dispersion Suspensions

Joanicot, Ajdari, Science, 2005


NanoNano--Pores: DNAPores: DNA--Positioning and Positioning and Pressure Control Pressure Control WinterhalterWinterhalter group, CF,...group, CF,...

PoreChannel 2

Channel 1

PoreChannel 2

Channel 1


Micro Patterning for Cell AssaysMicro Patterning for Cell AssaysPiel Piel & & BornensBornens, , Institut Institut CurieCurie


Cells Adapt Strongly to External ConditionsCells Adapt Strongly to External Conditions


Cell Assay on Patterned SurfacesCell Assay on Patterned Surfaces


Cells on LCells on L--Substate Substate Close to Migrating CellClose to Migrating Cell


Cell Cycle Becomes ReproducibleCell Cycle Becomes Reproducible


22--Dimensional GradientsDimensional Gradients

Hung, Lee, et al. 2004


SymmetrySymmetry


Magnetic Bead Micro ReactorsMagnetic Bead Micro Reactors

Slovacova Futterer et al., Lab Chip 2005


Fast Protein Digestion in Fast Protein Digestion in MicrochannelsMicrochannels


Rapid Cancer DiagnosisRapid Cancer Diagnosis


Nano Nano TransportTransport

Hess et al. 2006

Introduction to Microfluidics4 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006 Why...

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