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Instituto Superior Técnico
2009/2010




                                    Engenharia de Células e Tecidos

                             By: David Conceição nº64405 MBioNano
                                 Johannes Österreicher nº68694 MEng.Quimica
1. What is microfluidics?


   2. Micro –fabrication process


              3. Applications

                 3.1 State of the art


                   3.2 Case studies


              4. Conclusions

Microfluidics for cell culture
1. What is microfluidics?


 The science and technology of systems that process or manipulate
 small (sub-microliter-scale: nl, pl, fl and below) amounts of
 fluids,using channels with dimensions of tens to hundreds of
 micrometres or below.




                         I’m too big for
                            this stuff!




                                                     Microfluidic Chemostat
Microfluidics for cell culture
- Low sample consumption
       - Fast analysis times
       - High throughput screening
       - Possibility of efficient data
       - Allow single-use devices
       - Incorporation of sample preparation and
        product purification elements
       - Massively parallel microfabrication                   Laminar flow
       - Ease of making different designs
       - Replicate essays in same device possible


                    The behaviour of fluids in the micro-scale is very different than in
                    the macro-scale


Microfluidics for cell culture
Turbulent flow
                   Mixing


 Fluid behaviour depends on several parameters


       -Reynolds number
       - Navier Stokes equations
       - Knudsen number
       - Peclet number




Microfluidics for cell culture
Once upon a time…

   Started in the 1980s in microanalytical chemistry and later in
   biodefence, molecular biology and microelectronics

              - gas-phase chromatography (GPC), high-pressure liquid
              chromatography (HPLC) and capillary electrophoresis
              (CE);

              - detectors for chemical and biological threats;

              - high-throughput DNA sequencing;

              - microelectromechanical systems (MEMS)




Microfluidics for cell culture
1. What is microfluidics?


              2. Micro –fabrication method


              3. Applications

                 3.1 State of the art


                   3.2 Case studies


              4. Conclusions

Microfluidics for cell culture
1. 2. Micro-fabrication process


                                                 UV light
                                                Photomask
                                                photoresist


                                               Microchannel-
                                              patterned mold


                                             Pouring PDMS on
                                              the patterned
                                                   mold

                                               PDMS curing,
                                                cutting, and
                                                  bonding
Microfluidics for cell culture
The ‘plumbing’
        Microvalves

        Micropumps

        Microchannels




Microfluidics for cell culture
1. What is microfluidics?


              2. Micro –fabrication method


                         3. Applications

                 3.1 State of the art


                   3.2 Case studies


              4. Conclusions

Microfluidics for cell culture
1. 2.       3. Applications

      Microfluidics for cell culture

Macro-scale systems are completely different from the
in vivo-environment : different behaviour of cells!


Microfluidics can mimic the in vivo-
environment of cells (channels, tissues,
membranes etc.)


   Example: Human circulatory system to feed
   the cells and remove waste products




Microfluidics for cell culture
Bioanalysis




Microfluidics for cell culture
Cell- on a chip




Microfluidics for cell culture
1. 2. 3.   3.1 State of the art




Microfluidics for cell culture
Schematic suggesting the assembly of
                                       modules in a microfluidic chip.




Schematic diagram of reversible sealing of
microfluidic arrays onto microwell patterned
substrates to fabricate multiphenotype cell
arrays.

Microfluidics for cell culture
1. 2. 3. 3.1   3.2 Case studies

Two cell populations with intercellular
           communication
                                                 System design




Microfluidics for cell culture
Demonstration with coloured water (flushing, gradients, etc




Microfluidics for cell culture
Experiments with living cells:

                  N9 Microglia

                  SY5Y Neuroblastoma

                  To study intracellular communication involved in
                  brain diseases




Microfluidics for cell culture
Integrated microfluidic cell culture
             and lysis on a chip




Microfluidics for cell culture
Cultured HeLa, MCF-7, Jurkat, and CHO-K1 cells for
       up to five days and subsequently lysing the cells
       without adding lysing reagents


                                 Cell loading




                                                Images of breast cancer cells (MCF-7)
                                                cultured inside a device. After 72 h, the cells
                                                have formed spheroids. Scale bar is 75 mm.

Microfluidics for cell culture
Dynamics of cell lysis by electrochemically
              generated hydroxide


Microfluidics for cell culture
1. What is microfluidics?


              2. Micro –fabrication method


                         3. Applications

                 3.1 State of the art


                   3.2 Case studies


                         4. Conclusions

Microfluidics for cell culture
1. 2. 3. 3.1 3.2                      4. Conclusions


         Microfluidic devices can be a start ing point for lab on a chip,
         joining multiple operations.

         It can simulate the in vivo conditions.


                                            Microfluidic platforms are very useful
                                            alternatives for future cell biology studies
                                            and cell-based assays.
  Most culture systems do not utilize a variety
              of cellular analyses
                                                                            Cross
High surface-to-volume ratio of microchannels enhances the              contamination
adsorption of molecules onto channels walls, reducing the
effective concentration of reagents

                                             Dificulty controlling fluid behaviour
Microfluidics for cell culture
Bibliography
   - Bruzewicz, D., McGuigan, A. & Whitesides, G. Fabrication of a modular tissue construct in a microfluidic
   chip Lab on a Chip, Royal Society of Chemistry, 2008, Vol. 8(5), pp. 663-671
   - Khademhosseini, A., Yeh, J., Eng, G., Karp, J., Kaji, H., Borenstein, J., Farokhzad, O. & Langer, R. Cell
   docking inside microwells within reversibly sealed microfluidic channels for fabricating multiphenotype cell
   arrays Lab on a Chip, Royal Society of Chemistry, 2005, Vol. 5(12), pp. 1380-1386

   - Lovchik, R., Tonna, N., Bianco, F., Matteoli, M. & Delamarche, E. A microfluidic device for depositing and
   addressing two cell populations with intercellular population communication capability Biomedical
   Microdevices, Springer, 2010, Vol. 12, pp. 275–-282

   - Nevill, J., Cooper, R., Dueck, M., Breslauer, D. & Lee, L. Integrated microfluidic cell culture and lysis on a
   chip Lab on a Chip, Royal Society of Chemistry, 2007, Vol. 7(12), pp. 1689-1695

   - Wheeler, A., Throndset, W., Whelan, R., Leach, A., Zare, R., Liao, Y., Farrell, K., Manger, I., Daridon, A. &
   others Microfluidic device for single-cell analysis Analytical Chemistry, ACS Publications, 2003, Vol. 75(14),
   pp. 3581-3586

   - Whitesides, G. The origins and the future of microfluidics Nature, Nature Publishing Group, 2006, Vol.
   442(7101), pp. 368-373

   - Young, E. & Beebe, D. Fundamentals of microfluidic cell culture in controlled microenvironments Chemical
   Society Reviews, Royal Society of Chemistry, 2010, Vol. 39(3), pp. 1036-1048

Microfluidics for cell culture
The end




                                       Any questions?

Microfluidics for cell culture

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Microfluidics Presentation

  • 1. Instituto Superior Técnico 2009/2010 Engenharia de Células e Tecidos By: David Conceição nº64405 MBioNano Johannes Österreicher nº68694 MEng.Quimica
  • 2. 1. What is microfluidics? 2. Micro –fabrication process 3. Applications 3.1 State of the art 3.2 Case studies 4. Conclusions Microfluidics for cell culture
  • 3. 1. What is microfluidics? The science and technology of systems that process or manipulate small (sub-microliter-scale: nl, pl, fl and below) amounts of fluids,using channels with dimensions of tens to hundreds of micrometres or below. I’m too big for this stuff! Microfluidic Chemostat Microfluidics for cell culture
  • 4. - Low sample consumption - Fast analysis times - High throughput screening - Possibility of efficient data - Allow single-use devices - Incorporation of sample preparation and product purification elements - Massively parallel microfabrication Laminar flow - Ease of making different designs - Replicate essays in same device possible The behaviour of fluids in the micro-scale is very different than in the macro-scale Microfluidics for cell culture
  • 5. Turbulent flow Mixing Fluid behaviour depends on several parameters -Reynolds number - Navier Stokes equations - Knudsen number - Peclet number Microfluidics for cell culture
  • 6. Once upon a time… Started in the 1980s in microanalytical chemistry and later in biodefence, molecular biology and microelectronics - gas-phase chromatography (GPC), high-pressure liquid chromatography (HPLC) and capillary electrophoresis (CE); - detectors for chemical and biological threats; - high-throughput DNA sequencing; - microelectromechanical systems (MEMS) Microfluidics for cell culture
  • 7. 1. What is microfluidics? 2. Micro –fabrication method 3. Applications 3.1 State of the art 3.2 Case studies 4. Conclusions Microfluidics for cell culture
  • 8. 1. 2. Micro-fabrication process UV light Photomask photoresist Microchannel- patterned mold Pouring PDMS on the patterned mold PDMS curing, cutting, and bonding Microfluidics for cell culture
  • 9. The ‘plumbing’ Microvalves Micropumps Microchannels Microfluidics for cell culture
  • 10. 1. What is microfluidics? 2. Micro –fabrication method 3. Applications 3.1 State of the art 3.2 Case studies 4. Conclusions Microfluidics for cell culture
  • 11. 1. 2. 3. Applications Microfluidics for cell culture Macro-scale systems are completely different from the in vivo-environment : different behaviour of cells! Microfluidics can mimic the in vivo- environment of cells (channels, tissues, membranes etc.) Example: Human circulatory system to feed the cells and remove waste products Microfluidics for cell culture
  • 13. Cell- on a chip Microfluidics for cell culture
  • 14. 1. 2. 3. 3.1 State of the art Microfluidics for cell culture
  • 15. Schematic suggesting the assembly of modules in a microfluidic chip. Schematic diagram of reversible sealing of microfluidic arrays onto microwell patterned substrates to fabricate multiphenotype cell arrays. Microfluidics for cell culture
  • 16. 1. 2. 3. 3.1 3.2 Case studies Two cell populations with intercellular communication System design Microfluidics for cell culture
  • 17. Demonstration with coloured water (flushing, gradients, etc Microfluidics for cell culture
  • 18. Experiments with living cells: N9 Microglia SY5Y Neuroblastoma To study intracellular communication involved in brain diseases Microfluidics for cell culture
  • 19. Integrated microfluidic cell culture and lysis on a chip Microfluidics for cell culture
  • 20. Cultured HeLa, MCF-7, Jurkat, and CHO-K1 cells for up to five days and subsequently lysing the cells without adding lysing reagents Cell loading Images of breast cancer cells (MCF-7) cultured inside a device. After 72 h, the cells have formed spheroids. Scale bar is 75 mm. Microfluidics for cell culture
  • 21. Dynamics of cell lysis by electrochemically generated hydroxide Microfluidics for cell culture
  • 22. 1. What is microfluidics? 2. Micro –fabrication method 3. Applications 3.1 State of the art 3.2 Case studies 4. Conclusions Microfluidics for cell culture
  • 23. 1. 2. 3. 3.1 3.2 4. Conclusions Microfluidic devices can be a start ing point for lab on a chip, joining multiple operations. It can simulate the in vivo conditions. Microfluidic platforms are very useful alternatives for future cell biology studies and cell-based assays. Most culture systems do not utilize a variety of cellular analyses Cross High surface-to-volume ratio of microchannels enhances the contamination adsorption of molecules onto channels walls, reducing the effective concentration of reagents Dificulty controlling fluid behaviour Microfluidics for cell culture
  • 24. Bibliography - Bruzewicz, D., McGuigan, A. & Whitesides, G. Fabrication of a modular tissue construct in a microfluidic chip Lab on a Chip, Royal Society of Chemistry, 2008, Vol. 8(5), pp. 663-671 - Khademhosseini, A., Yeh, J., Eng, G., Karp, J., Kaji, H., Borenstein, J., Farokhzad, O. & Langer, R. Cell docking inside microwells within reversibly sealed microfluidic channels for fabricating multiphenotype cell arrays Lab on a Chip, Royal Society of Chemistry, 2005, Vol. 5(12), pp. 1380-1386 - Lovchik, R., Tonna, N., Bianco, F., Matteoli, M. & Delamarche, E. A microfluidic device for depositing and addressing two cell populations with intercellular population communication capability Biomedical Microdevices, Springer, 2010, Vol. 12, pp. 275–-282 - Nevill, J., Cooper, R., Dueck, M., Breslauer, D. & Lee, L. Integrated microfluidic cell culture and lysis on a chip Lab on a Chip, Royal Society of Chemistry, 2007, Vol. 7(12), pp. 1689-1695 - Wheeler, A., Throndset, W., Whelan, R., Leach, A., Zare, R., Liao, Y., Farrell, K., Manger, I., Daridon, A. & others Microfluidic device for single-cell analysis Analytical Chemistry, ACS Publications, 2003, Vol. 75(14), pp. 3581-3586 - Whitesides, G. The origins and the future of microfluidics Nature, Nature Publishing Group, 2006, Vol. 442(7101), pp. 368-373 - Young, E. & Beebe, D. Fundamentals of microfluidic cell culture in controlled microenvironments Chemical Society Reviews, Royal Society of Chemistry, 2010, Vol. 39(3), pp. 1036-1048 Microfluidics for cell culture
  • 25. The end Any questions? Microfluidics for cell culture