Introduction to Microfluidics. C. Fütterer, Institut Curie & Fluigent SA, Paris

Transcription

1 Introduction to Microfluidics C. Fütterer, Institut Curie & Fluigent SA, Paris

2 Miniaturisation & Integration

3 Micro-Pipettes Problems: Minimal volume: 1μl Samples unprotected against evaporation & contamination (dust) Slow and tedious operation Manipulation errors Difficult automation through expensive robots 3 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

4 Why Traditional Instruments are Unsufficient Complexity: genes and more than 10 6 proteins in human body. Spatio-temporal dynamics of cellular/molecular processes. Big volumes: low speed of thermal and molecular diffusion and reactions. 4 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

5 Size of Biological Objects pl nl μl ml? 5 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

6 Solution: Microfluidics Come Closer to Cells and Molecules! Institut Curie Smallest fluid quantities (single cell) High throughput (statistics) Faster physico-chemical processes (diffusion more efficient) Towards quantitative biology! 6 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

7 Pioneer Publication Electronic Chips 7 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

8 Chips are not Chips... Electronic Chips Microfluidic Chips DNA-Chips Institut Curie 8 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

9 Many Advantages 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. 9 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

10 Microfluidics is Not New - Arteria and venes in animals - Capillaries in plants 10 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

11 How to Make it: Microfabrication

12 Photolithography Mask: PROTOCOL Spincoat photosensitive solution, Dry Put mask on top Expose to UV Wash Heat Master 12 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

13 Rapid Prototyping: Principle Chip-Fabrication Master Surface-Patterning by μ Contact Printing Chip (PDMS) Master 13 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

14 Rapid Prototyping: Mask and Mould Chip-Fabrication Chip (PDMS) 14 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

15 Rapid Prototyping: Mould Chip-Fabrication Chip (PDMS) 15 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

16 Mould: Vials Reproducible and clean vials Polymer Chip (PDMS) Master Air Microstructure 16 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

17 Rapid Prototyping: Master I Chip-Fabrication Chip (PDMS) Much more stable! Pallandre, Fütterer et al., Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

18 Rapid Prototyping: Master II Chip-Fabrication Chip (PDMS) 18 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

19 Rapid Prototyping: : Polyester Master Chip (PDMS) 19 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

20 Taylored Channel Systems, Integration Quake et al Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

21 Flow Control

22 Micro-Fluidics Fluidics: Defined Flow Profile Laminar profile 22 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

23 Difficulty: Flow Control through Interfacing (Contraction) Human scale 1cm 10μm Molecules and Cells Ratio: 10 6!!! 23 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

24 Flow Control: Syringe Pumps Syringe pumps: Long response time Strong hysteresis Limited volume Stopped-flow/flow inversion difficult Slow Bubbles Mechanical drive Bending of tubes Atmospheric Pressure Elasticity 24 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

25 Flow Control: Electroosmotic Pumps Reproducibility problems with biological fluids Time consuming preparation 25 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

26 Flow Control: AC-Electroosmotic Pumps Reproducibility problems with biological fluids Time consuming preparation Electrodes are consumed mutas Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

27 Flow Control: Microfabricated Pumps Pulsed flow Complex Microfabrication Objects are squeezed (Cells!) Pressure Quake et al Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

28 Flow Control Peristaltic pumps Pulses Long time constant Objects are squeezed Tubes have to be exchanged 28 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

29 Capillary Forces or Centrifugal Systems Blood test Separation of blood plasma (Zengerle et al. IMTEK) 29 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

30 Pumping through Moving Objects Optical tweezer Optical tweezer Leach et al. Lab Chip Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

31 New Principle for Microfluidics Flow Control Semi-open pneumatic pressure control Pneumatic Wheatstone bridge Microchannel Calibrated leaks 31 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

32 New Principle for Microfluidics Flow Control 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 Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

33 Vision of the Lab today tomorrow Microfluidic Chip: 33 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

34 MicroFluidic Control System Start-up Fluigent Industrial prototype 20cm Pressure output Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

35 Very Fast Response in Microchannel single T4 DNA molecule playing with sliding knob 35 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

36 Applications

37 Low Dispersion Suspensions Joanicot, Ajdari, Science, Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

38 Nano-Pores: DNA-Positioning and Pressure Control Winterhalter group, CF,... Channel 1 Channel 2 Pore Channel 1 Channel 2 Pore 38 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

39 Micro Patterning for Cell Assays Piel & Bornens, Institut Curie 39 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

40 Cells Adapt Strongly to External Conditions 40 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

41 Cell Assay on Patterned Surfaces 41 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

42 Cells on L-Substate L Close to Migrating Cell 42 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

43 Cell Cycle Becomes Reproducible 43 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

44 2-Dimensional Gradients Hung, Lee, et al Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

45 Symmetry 45 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

46 Magnetic Bead Micro Reactors Slovacova Futterer et al., Lab Chip Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

47 Fast Protein Digestion in Microchannels 47 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

48 Rapid Cancer Diagnosis 48 - Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006

49 Nano Transport Hess et al Microfluidics & Magnetic Beads - C. Fütterer - 23/09/2006