Advertisement

Design and characterization of a hydrodynamically confined microflow device for applying controlled loads to investigate single-cell mechanics

  • Kevin V. Christ
  • Choongbae ParkEmail author
  • Kristyn S. Masters
  • Kevin T. TurnerEmail author
Research Paper
  • 142 Downloads

Abstract

Single-cell mechanics measurements are crucial in understanding mechanotransduction and cellular properties, such as adhesion and stiffness. Here, we present a microfluidic probe device that can generate controlled hydrodynamic loads on single cells in an open cell culture environment. The device is optimized to produce uniform stresses across the area of a cell for cell adhesion measurements. Microfluidic probe (MFP) devices that can be used to create hydrodynamically confined flows (HCMs) have emerged as a unique device for selectively treating cells and surfaces. Typical MFP devices generate complex-shaped flows and non-uniform hydrodynamic loads on the surface beneath the device. We have used computational fluid dynamics to optimize the port geometry of the MFP device to generate an HCM with uniform shear stresses in a region beneath the device. The devices were fabricated from a combination of silicon and PDMS and characterized through flow experiments above a polyacrylamide gel seeded with fluorescent beads. Bead displacements were measured as a function of flow conditions and general agreement with the model was obtained. Finally, we have used the devices to characterize the adhesion strength of patterned fibroblast cells adhered to a collagen-coated substrate. The results presented establish a design for an MFP device that can apply controlled mechanical forces to cells in open liquid environments.

Keywords

Microfluidic probe Hydrodynamically confined flows Computational fluid dynamics-based design Cell mechanics 

Notes

Acknowledgements

We acknowledge financial support from a 3M fellowship, the Wisconsin Alumni Research Foundation, and NSF award 0832802 at the University of Pennsylvania. Finally, we thank Dr. David S. Grierson for assistance in performing the AFM measurements on the polyacrylamide gels. K.S.M. acknowledges support from NIH R01 HL093281.

Compliance with ethical standards

Conflict of interest

There are no conflicts to declare.

References

  1. Albro MB, Li R, Banerjee RE, Hung CT, Ateshian GA (2010) Validation of theoretical framework explaining active solute uptake in dynamically loaded porous media. J Biomech 43:2267–2273.  https://doi.org/10.1016/j.jbiomech.2010.04.041 CrossRefGoogle Scholar
  2. Anselme K (2000) Osteoblast adhesion on biomaterials. Biomaterials 21:667–681CrossRefGoogle Scholar
  3. Bao G, Suresh S (2003) Cell and molecular mechanics of biological materials. Nat Mater 2:715–725.  https://doi.org/10.1038/Nmat1001 CrossRefGoogle Scholar
  4. Beningo KA, Lo CM, Wang YL (2002) Flexible polyacrylamide substrata for the analysis of mechanical interactions at cell-substratum adhesions. Method Cell Biol 69:325–339CrossRefGoogle Scholar
  5. Bhana B et al (2010) Influence of substrate stiffness on the phenotype of heart cells. Biotechnol Bioeng 105:1148–1160.  https://doi.org/10.1002/bit.22647 CrossRefGoogle Scholar
  6. Bhattacharya S, Datta A, Berg JM, Gangopadhyay S (2005) Studies on surface wettability of poly(dimethyl) siloxane (PDMS) and glass under oxygen-plasma treatment and correlation with bond strength. J Microelectromech Syst 14:590–597.  https://doi.org/10.1109/Jmems.2005.844746 CrossRefGoogle Scholar
  7. Brock A, Chang E, Ho CC, LeDuc P, Jiang XY, Whitesides GM, Ingber DE (2003) Geometric determinants of directional cell motility revealed using microcontact printing. Langmuir 19:1611–1617 doi.  https://doi.org/10.1021/La026394k CrossRefGoogle Scholar
  8. Cavallaro U, Christofori G (2004) Cell adhesion and signalling by cadherins and Ig-CAMs in cancer. Nat Rev Cancer 4:118–132 doi.  https://doi.org/10.1038/Nrc1276 CrossRefGoogle Scholar
  9. Chen CS (2008) Mechanotransduction—a field pulling together? J Cell Sci 121:3285–3292.  https://doi.org/10.1242/jcs.023507 CrossRefGoogle Scholar
  10. Christ KV, Turner KT (2010) Methods to measure the strength of cell adhesion to substrates. J Adhes Sci Technol 24:2027–2058CrossRefGoogle Scholar
  11. Christ KV, Turner KT (2011) Design of hydrodynamically confined microfluidics: controlling flow envelope and pressure. Lab Chip 11:1491–1501.  https://doi.org/10.1039/c0lc00416b CrossRefGoogle Scholar
  12. Christ KV, Williamson KB, Masters KS, Turner KT (2010) Measurement of single-cell adhesion strength using a microfluidic assay. Biomed Microdevice 12:443–455CrossRefGoogle Scholar
  13. Cross SE, Jin YS, Rao J, Gimzewski JK (2007) Nanomechanical analysis of cells from cancer patients. Nat Nanotechnol 2:780–783.  https://doi.org/10.1038/nnano.2007.388 CrossRefGoogle Scholar
  14. Delamarche E, Kaigala GV (eds) (2018) Open-space microfluidics: concepts, implementations, applications. Wiley, New YorkGoogle Scholar
  15. Desgrosellier JS, Cheresh DA (2010) Integrins in cancer: biological implications and therapeutic opportunities. Nat Rev Cancer 10:9–22.  https://doi.org/10.1038/nrc2748 CrossRefGoogle Scholar
  16. Desmaele D, Boukallel M, Regnier S (2011) Actuation means for the mechanical stimulation of living cells via microelectromechanical systems: a critical review. J Biomech 44:1433–1446.  https://doi.org/10.1016/j.jbiomech.2011.02.085 CrossRefGoogle Scholar
  17. Engler AJ, Richert L, Wong JY, Picart C, Discher DE (2004) Surface probe measurements of the elasticity of sectioned tissue, thin, gels and polyelectrolyte multilayer films: correlations between substrate stiffness and cell adhesion. Surf Sci 570:142–154CrossRefGoogle Scholar
  18. Gallant ND, Michael KE, Garcia AJ (2005) Cell adhesion strengthening: contributions of adhesive area, integrin binding and focal adhesion assembly. Mol Biol Cell 16:4329–4340.  https://doi.org/10.1091/mbc.E05-02-0170 CrossRefGoogle Scholar
  19. Garcia AJ, Gallant ND (2003) Stick and grip—measurement systems and quantitative analyses of integrin-mediated cell adhesion strength. Cell Biochem Biophys 39:61–73CrossRefGoogle Scholar
  20. Gaver DP, Kute SM (1998) A theoretical model study of the influence of fluid stresses on a cell adhering to a microchannel wall. Biophys J 75:721–733CrossRefGoogle Scholar
  21. Griffin MA, Engler AJ, Barber TA, Healy KE, Sweeney HL, Discher DE (2004) Patterning, prestress, and peeling dynamics of myocytes. Biophys J 86:1209–1222CrossRefGoogle Scholar
  22. Guilak F, Cohen DM, Estes BT, Gimble JM, Liedtke W, Chen CS (2009) Control of stem cell fate by physical interactions with the extracellular matrix. Cell Stem Cell 5:17–26.  https://doi.org/10.1016/j.stem.2009.06.016 CrossRefGoogle Scholar
  23. Hamill OP, Martinac B (2001) Molecular basis of mechanotransduction in living cells. Physiol Rev 81:685–740CrossRefGoogle Scholar
  24. Hersel U, Dahmen C, Kessler H (2003) RGD modified polymers: biomaterials for stimulated cell adhesion and beyond. Biomaterials 24:4385–4415.  https://doi.org/10.1016/S0142-9612(03)00343-0 CrossRefGoogle Scholar
  25. Hu YH, Suo ZG (2012) Viscoelasticity and poroelasticity in elastomeric gels. Acta Mechanica Solida Sinica 25:441–458  https://doi.org/10.1016/s0894-9166(12)60039-1 CrossRefGoogle Scholar
  26. Huang S, Ingber DE (1999) The structural and mechanical complexity of cell-growth control. Nat Cell Biol 1:E131–E138CrossRefGoogle Scholar
  27. Ingber DE (2006) Cellular mechanotransduction: putting all the pieces together again. FASEB J 20:811–827.  https://doi.org/10.1096/fj.05-5424rev CrossRefGoogle Scholar
  28. Juncker D, Schmid H, Delamarche E (2005) Multipurpose microfluidic probe. Nat Mater 4:622–628.  https://doi.org/10.1038/Nmat1435 CrossRefGoogle Scholar
  29. Kaigala GV, Lovchik RD, Drechsler U, Delamarche E (2011) A vertical microfluidic probe. Langmuir 27:5686–5693  https://doi.org/10.1021/la2003639 CrossRefGoogle Scholar
  30. Khalili AA, Ahmad MR (2015) A review of cell adhesion studies for biomedical and biological applications. Int J Mol Sci 16:18149–18184.  https://doi.org/10.3390/ijms160818149 CrossRefGoogle Scholar
  31. Korson L, Drost-Hansen W, Millero FJ (1969) Viscosity of water at various temperatures. J Phys Chem 73:34–39.  https://doi.org/10.1021/j100721a006 CrossRefGoogle Scholar
  32. Lauffenburger DA, Horwitz AF (1996) Cell migration: a physically integrated molecular process. Cell 84:359–369CrossRefGoogle Scholar
  33. Lovchik RD, Kaigala GV, Georgiadis M, Delamarche E (2012) Micro-immunohistochemistry using a microfluidic probe. Lab Chip 12:1040–1043.  https://doi.org/10.1039/c2lc21016a CrossRefGoogle Scholar
  34. Lu H, Koo LY, Wang WM, Lauffenburger DA, Griffith LG, Jensen KF (2004) Microfluidic shear devices for quantitative analysis of cell adhesion. Anal Chem 76:5257–5264.  https://doi.org/10.1021/Ac049837t CrossRefGoogle Scholar
  35. Qasaimeh MA, Gervais T, Juncker D (2011) Microfluidic quadrupole and floating concentration gradient. Nat Commun 2:464.  https://doi.org/10.1038/ncomms1471 CrossRefGoogle Scholar
  36. Queval A, Ghattamaneni NR, Perrault CM, Gill R, Mirzaei M, McKinney RA, Juncker D (2010) Chamber and microfluidic probe for microperfusion of organotypic brain slices. Lab Chip 10:326–334.  https://doi.org/10.1039/B916669f CrossRefGoogle Scholar
  37. Quinn TM (2013) Flow-induced deformation of poroelastic tissues and gels: a new perspective on equilibrium pressure-flow-thickness relations. J Biomech Eng Trans Asme.  https://doi.org/10.1115/1.4023095 CrossRefGoogle Scholar
  38. Rajagopalan J, Saif MT (2011) MEMS sensors and microsystems for cell mechanobiology. J Micromech Microeng 21:54002–54012.  https://doi.org/10.1088/0960-1317/21/5/054002 CrossRefGoogle Scholar
  39. Safavieh M, Qasaimeh MA, Vakil A, Juncker D, Gervais T (2015) Two-aperture microfluidic probes as flow dipoles: theory and applications. Sci Rep 5:11943CrossRefGoogle Scholar
  40. Schwartz MA, DeSimone DW (2008) Cell adhesion receptors in mechanotransduction. Curr Opin Cell Biol 20:551–556.  https://doi.org/10.1016/j.ceb.2008.05.005 CrossRefGoogle Scholar
  41. Takayama S, McDonald JC, Ostuni E, Liang MN, Kenis PJA, Ismagilov RF, Whitesides GM (1999) Patterning cells and their environments using multiple laminar fluid flows in capillary networks. Proc Natl Acad Sci USA 96:5545–5548CrossRefGoogle Scholar
  42. Takigawa T, Morino Y, Urayama K, Masuda T (1996) Poisson’s ratio of polyacrylamide (PAAm) gels. Polym Gels Netw 4:1–5CrossRefGoogle Scholar
  43. Thoumine O, Cardoso O, Meister J-J (1999) Changes in the mechanical properties of fibroblasts during spreading: a micromanipulation study. Eur Biophys J 28:222–234CrossRefGoogle Scholar
  44. Van Vliet KJ, Bao G, Suresh S (2003) The biomechanics toolbox: experimental approaches for living cells and biomolecules. Acta Mater 51:5881–5905.  https://doi.org/10.1016/j.actamat.2003.09.001 CrossRefGoogle Scholar
  45. Xia Y, Whitesides GM (1998) Soft lithography. Angewandte Chemie-International Edition 37:551–575CrossRefGoogle Scholar
  46. Yamamoto A, Mishima S, Maruyama N, Sumita M (2000) Quantitative evaluation of cell attachment to glass, polystyrene, and fibronectin- or collagen-coated polystyrene by measurement of cell adhesive shear force and cell detachment energy. J Biomed Mater Res 50:114–124CrossRefGoogle Scholar
  47. COMSOL (2019) COMSOL Multiphysics reference manual, version 4.4. COMSOL, IncGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Mechanical EngineeringUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Department of Mechanical Engineering and Applied MechanicsUniversity of PennsylvaniaPhiladelphiaUSA
  3. 3.Department of Mechanical and Industrial EngineeringTexas A&M University-KingsvilleKingsvilleUSA
  4. 4.Department of Biomedical EngineeringUniversity of Wisconsin-MadisonMadisonUSA
  5. 5.Aerospace Advanced Technology, Honeywell InternationalPlymouthUSA

Personalised recommendations