Microfluidics A new technology. A review Marie C Béné Nantes France

Transcription

1 Microfluidics A new technology A review Marie C Béné Nantes France

2 Nothing to disclose

3 CELL COUNTING

4 CELL TYPING

5 TOWARDS MICROFLUIDICS Lab on a chip or Lab-on-chip : LOC LOC represents the next generation of analytical laboratories that have been miniaturized to the size of a matchbox, and represent one of the most groundbreaking offshoots of nanotechnology Donald Wlodkowic and Zbigniew Darzynkiewicz Methods Cell Biol 2011 Since April 2017 issue IF 5.59

6 TOWARDS MICROFLUIDICS Pubmed search April 2017 Microfluidics : ~9000 Microfluidics and cell counting : 136 Microfluidics and flow cytometry : 327 Microfluidics and Lab-on-chip : 98 Lab on chip : 9146 Lab on chip and cytometry : 327, 36 reviews

7 APPLICATIONS OF MICROFLUIDICS Immunoassays Separation of proteins Separation of nucleic acids Cell sorting/counting : bacteria, cancer cells, leukocytes Flow cytometry Flow cytometry and cell sorting Magnetic cell sorting Cell biology Drug testing

8 CELL COUNTING AND CELL TYPING CURRENT Samples Count : ml Flow : 50/100µL 1/2 ml Reagents Lysis Buffer Stains Fluidics Sheath Optics Waste disposal Electronics MICROFLUIDICS Smaller samples Less reagents Fluidics or not! Optics Waste disposal Electronics

9 DIFFERENT PHYSICS Microfluidics versus macrofluidics Microchannels of µm Influence of geometry, fluid viscosity and average flow rates Reynolds parameter : Re=vlr/µ v= velocity (m/s) l = cross sectional dimension (m) R = density of the fluid (1000Kg/m 3 for water) µ = viscosity of the fluid (water 10-3 kg/m.s) Typical <100 µm channels and low flow 1cm/s : Re~1 : laminar flow Re>~2000 : turbulent flow Laminar flow for multiple fluid streams Mixing by diffusion Mixing by flow motion Turbulent flows Mixing of the streams : built in devices and/or pressure

10 HOW TO MAKE MICROFLUIDICS DEVICES

11 MICROFLUIDIC DEVICES Components Microchannels Microwells Waste and sorting collectors Accessories Pumps Sensors Optics Lasers and Photomultipliers Signal collectors Materials Glass Silicone Plastics Polydimethylsiloxane : PMDS

12 PDMS Inexpensive Flexible Optically transparent Water proof Non toxic Permeable to gases Easily prototyped Easily bonded to other surfaces

13 MICROFLUIDICS DEVICES Soft lithography Computer assisted design of the pattern : generation of a mask Use mask on photoresist material + UV light Generation of the maze Solidification through UV light Disruption through UV light Washes : pods, wells, channels Obtention of a master Moulding of PDMS (x.. up to 100+) Sealing of PDMS on a surface Use of 3D printers

14 SORTING DEVICES Passive devices Indented, protruded or curved sidewalls in microchannels Inertial focusing, spirals, chevrons, pods Geometric patterns Transient binding to adhesion molecules Active devices «Classical» fluorescence cell sorting Generation of single-cell droplets «reaction vessels» Magnets Acoustic fields

15 EXAMPLES OF MICROFLUDICS

16 Deterministic lateral displacement DLD Small cells move with the convective flow; larger cells move in the direction given by the pods Shields CW et al. Lab Chip 2015

17 Another example of DLD Civin CA et al. Cytometry part A, 2016

18 Inertial microfluidics in a spiral From Kuntaegowdanahalli et al. In Shields CW et al. Lab Chip 2015

19 Impedance flow cytometric microfluidics Chen et al Int J Mol Sci. 2015

20 Circulating tumor cells with magnetic beads From Ozkumar et al. In Shields CW et al. Lab Chip 2015

21 A hand-held flow cytometer Grafton MMG et al. SPIE proceeedings 2011

22 Fishman for CTC Watanabe M et al. J Transl Med 2014

23 Gigasort Hulspas R et al. Cytotherapy 2014

24 Acoustophoric microchip Augustsson P et al. In Piyasena ME and Graves SW Lab Chip 2014

25 Sony SH800

26 Picovitro plates for cell culture Lindström S In Wlodkowic D and Darzynkiewicz Z Methods Cell Biol 2012

27 Antibody production in droplets Mazutis L et al. Nat Protocols 2014

28 Exosomes Kanwar SS et al. Lab chip 2014

29 SUMMARY OF THE PROPERTIES OF MICROFLUIDICS Shields CW et al. Cytometry part B, 2017 Sorting performances nearly comparable to FACS and MACS devices Spatial (e.g., to a precision of 1 µm) and temporal control of cells Preserved cell function by obviating the need for high-pressure sheath fluids Multiplexed cell sorting capabilities Decreased unit cost i.e. <$5 per microfluidic cell Significantly decreased power consumption Potential disposability Ability to process unmodified biological fluids Improved portability Reduced reagent consumption Controlled conditions (temperature, gas, ph ) Commercially available?

30 A PROMISING FUTURE???

31 THANK YOU FOR YOUR ATTENTION