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.
Similar content being viewed by others
References
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
Anselme K (2000) Osteoblast adhesion on biomaterials. Biomaterials 21:667–681
Bao G, Suresh S (2003) Cell and molecular mechanics of biological materials. Nat Mater 2:715–725. https://doi.org/10.1038/Nmat1001
Beningo KA, Lo CM, Wang YL (2002) Flexible polyacrylamide substrata for the analysis of mechanical interactions at cell-substratum adhesions. Method Cell Biol 69:325–339
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
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
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
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
Chen CS (2008) Mechanotransduction—a field pulling together? J Cell Sci 121:3285–3292. https://doi.org/10.1242/jcs.023507
Christ KV, Turner KT (2010) Methods to measure the strength of cell adhesion to substrates. J Adhes Sci Technol 24:2027–2058
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
Christ KV, Williamson KB, Masters KS, Turner KT (2010) Measurement of single-cell adhesion strength using a microfluidic assay. Biomed Microdevice 12:443–455
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
Delamarche E, Kaigala GV (eds) (2018) Open-space microfluidics: concepts, implementations, applications. Wiley, New York
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
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
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 570:142–154
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
Garcia AJ, Gallant ND (2003) Stick and grip—measurement systems and quantitative analyses of integrin-mediated cell adhesion strength. Cell Biochem Biophys 39:61–73
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 75:721–733
Griffin MA, Engler AJ, Barber TA, Healy KE, Sweeney HL, Discher DE (2004) Patterning, prestress, and peeling dynamics of myocytes. Biophys J 86:1209–1222
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
Hamill OP, Martinac B (2001) Molecular basis of mechanotransduction in living cells. Physiol Rev 81:685–740
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
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
Huang S, Ingber DE (1999) The structural and mechanical complexity of cell-growth control. Nat Cell Biol 1:E131–E138
Ingber DE (2006) Cellular mechanotransduction: putting all the pieces together again. FASEB J 20:811–827. https://doi.org/10.1096/fj.05-5424rev
Juncker D, Schmid H, Delamarche E (2005) Multipurpose microfluidic probe. Nat Mater 4:622–628. https://doi.org/10.1038/Nmat1435
Kaigala GV, Lovchik RD, Drechsler U, Delamarche E (2011) A vertical microfluidic probe. Langmuir 27:5686–5693 https://doi.org/10.1021/la2003639
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
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
Lauffenburger DA, Horwitz AF (1996) Cell migration: a physically integrated molecular process. Cell 84:359–369
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
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
Qasaimeh MA, Gervais T, Juncker D (2011) Microfluidic quadrupole and floating concentration gradient. Nat Commun 2:464. https://doi.org/10.1038/ncomms1471
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
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
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
Safavieh M, Qasaimeh MA, Vakil A, Juncker D, Gervais T (2015) Two-aperture microfluidic probes as flow dipoles: theory and applications. Sci Rep 5:11943
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
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 96:5545–5548
Takigawa T, Morino Y, Urayama K, Masuda T (1996) Poisson’s ratio of polyacrylamide (PAAm) gels. Polym Gels Netw 4:1–5
Thoumine O, Cardoso O, Meister J-J (1999) Changes in the mechanical properties of fibroblasts during spreading: a micromanipulation study. Eur Biophys J 28:222–234
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
Xia Y, Whitesides GM (1998) Soft lithography. Angewandte Chemie-International Edition 37:551–575
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 50:114–124
COMSOL (2019) COMSOL Multiphysics reference manual, version 4.4. COMSOL, Inc
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.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
There are no conflicts to declare.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Christ, K.V., Park, C., Masters, K.S. et al. Design and characterization of a hydrodynamically confined microflow device for applying controlled loads to investigate single-cell mechanics. Microfluid Nanofluid 23, 49 (2019). https://doi.org/10.1007/s10404-019-2210-5
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10404-019-2210-5