Abstract
We present a method of measuring force response of a single living cell, attached to a substrate, in situ, by a functionalized microelectromechanical systems sensor that applies local deformation on the cell. The sensor is a single crystal silicon microcantilever beam with prescribed shape and geometry, and is coated by a thin layer of fibronectin. It is brought in contact with a cell to form adhesion cites, and is then moved by a piezoactuator to deform the cell locally. The force is transmitted from the adhesion site(s) on the cantilever to the sites on the substrate through the cytoskeleton. The interaction force between the cell and the cantilever is measured from the deformation of the cantilever and its spring constant, which can be obtained by several independent means. The force and the cell deformation can be 10 s of nano-Newtons and micrometers, respectively. We demonstrate the method using two families of force sensors with spring constants of 18 and 0.4 nN/μm. Several cells, endothelial and fibroblast, are deformed by tens of micrometers until the adhesion sites failed. Their force–deformation response shows strong linearity. Several possible mechanisms are discussed to explain the linear response. © 2003 Biomedical Engineering Society.
PAC2003: 8780Fe, 0710Pz, 8716Gj, 8585+j
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References
Block, S.Making light work with optical tweezers. Nature (London)360:493–495, 1992.
Bowen, W. R., N. Hilal, R. W. Lovitt, and C. J. Wright. Direct measurement of the force of adhesion of a single biological cell using an atomic force microscope. Colloids Surf., A136:231–234, 1998.
Breen, E. C.Mechanical strain increases type I collagen expression in pulmonary fibroblasts. J. Appl. Physiol.88:203–209, 2000.
Chen, C. S., M. Mrksich, S. Huang, G. M. Whitesides, and D. E. Ingber. Geometric control of cell life and death. Science276:1425–1428, 1997.
Chen, K.-D., Y.-S. Li, M. Kim, S. Li, S. Yuan, S. Chien, and J. Y.-J. Shyy. Mechanotransduction in response to shear stress. J. Biol. Chem.274:18393–18400, 1999.
Chesla, S. E., P. Selvaraj, and C. Zhu. Measuring two-dimensional receptor–ligand binding kinetics by micropipette. Biophys. J.75:1553–1572, 1998.
Chicurel, M. E., C. S. Chen, and D. E. Ingber. Cellular control lies in the balance of forces. Curr. Opin. Cell Biol.10:232–239, 1998.
Clough, R. W., and J. Penzien. Dynamics of Structures. New York: McGraw-Hill, 1986, pp 137.
Crouch, C. J., H. W. Fowler, and R. E. Spier. The adhesion of animal cells to surfaces: The measurement of critical surface shear stress permitting attachment or causing detachment. J. Chem. Technol. Biotechnol.35B:273–281, 1985.
Easty, G. C., D. M. Easty, and E. J. Ambrose. Studies of cellular adhesiveness. Exp. Cell Res.19:539–548, 1960.
Evans, E., D. Berk, and A. Leung. Detachment of agglutinin-bonded red blood cells, I. Forces to rupture molecular-point attachments. Biophys. J.59:838–848, 1991.
Evans, E., D. Berk, A. Leung, and N. Mohandas. Detachment of agglutinin-bonded red blood cells, II. Mechanical energies to separate large contact areas. Biophys. J.59:849–860, 1991.
Florin, E.-L., V. T. Moy, and H. E. Gaub. Adhesion forces between individual ligand–receptor pairs. Science264:415–417, 1994.
Forrester, J. V., and J. M. Lackie. Adhesion of neutrophil leucocytes under conditions of flow. J. Cell. Sci.70:93–110, 1984.
Heidemann, S. R., S. Kaech, R. E. Buxbaum, and A. Matus. Direct observations of the mechanical behaviors of the cytoskeleton in living fibroblasts. J. Cell Biol.145:109–122, 1999.
Helm, C. A., W. Knoll, and J. N. Israelachvili. Measurement of ligand–receptor interactions. Proc. Natl. Acad. Sci. U.S.A.88:8169–8173, 1991.
Horwitz, A. F.Integrins and health. Sci. Am.276:68–75, 1997.
Hwang, W. C., and R. E. Waugh. Energy of dissociation of lipid bilayer from the membrane skeleton of red blood cells. Biophys. J.72:2669–2678, 1997.
Ingber, D.How cells (might) sense microgravity. FASEB J.13:S3–S15, 1999.
Ingber, D. E.The architecture of life. Sci. Am.278(1):30–39, 1998.
Kaech, S., B. Ludin, and A. Matus. Cytoskeletal plasticity in cells expressing neuronal microtubule-associated proteins. Neuron17:1189–1199, 1996.
Karila-Cohen, D., and S. P. Gabriel. Aspirin and prevention of cardiovascular risk. Rev. Med. Interne1:35s-40s, 2000.
Kirshner, M., and T. Mitchison. Beyond self-assembly: From microtubules to morphogenesis. Cell45:329–342, 1986.
Lauffenburger, D. A., and A. F. Horwitz. Cell Migration: A physically integrated molecular process. Cell84:359–369, 1996.
Li, X., and R. B. Nicklas. Tension-sensitive kinetochore phosphorylation and the chromosome distribution checkpoint in praying mantid spermatocytes. J. Cell. Sci.110:537–545, 1997.
Markel, R., P. Nassoy, A. Leung, K. Ritchie, and E. Evans. Energy landscapes of receptor–ligand bonds explored with dynamic force spectroscopy. Nature (London)397:50–53, 1999.
Pickett-Heaps, J. D., A. Forer, and T. Spurck. Traction fiber: Towards a tensegral model of the spindle. Cell Motil. Cytoskeleton37:1–6, 1997.
Ra, H. J., C. Picart, H. Feng, H. L. Sweeney, and D. E. Discher. Muscle cell peeling from micropatterned collagen: Direct probing of focal and molecular properties of matrix adhesion. J. Cell. Sci.112:1425–1436, 1999.
Ruoslahti, E., and J. C. Reed. Anchorage dependence, integrins, and apoptosis. Cell77:477–478, 1994.
Sager, C., P. LeDuc, and T. Saif. adhesion studies of a single living bovine endothelial cell using MEMS sensor. Procedings of the 1st Annual International IEEE–EMBS Special Topic Conference on Microtechnologies in Medicine and Biology,Palais des Congres, Lyon, France, 12–14 October 2000, pp. 76–79.
Sager, C., T. Saif, and P. LeDuc. Adhesion Studies of Cells Using MEMS Sensors. Proceedings of the ASME Winter Annual Meeting, MEMS, 2000, Orlando, FL, 5–10 November 2000, Vol. 2, pp. 215–219.
Saif, M. T. A.On the capillary interaction between solid plates forming menisci on the surface of a liquid. J. Fluid Mech.473:321–347, 2002.
Saif, M. T. A., and N. C. MacDonald. A milli-Newton microloading device. Sens. Actuators A52:65–75, 1996.
Saif, M. T. A., and N. C. MacDonald. Planarity of large MEMS. J. Microelectromech. Syst.5:79–97, 1996.
Saif, M. T. A., and N. C. MacDonald. Measurement of forces and spring constants of micro instruments. Rev. Sci. Instrum.69:1410–1422, 1998.
Shao, J.-Y., and R. M. Hochmuth. Mechanical anchoring strength of L-selectin, β2 integrins, and CD 45 to neutrophil cytoskeleton and membrane. Biophys. J.77:587–596, 1999.
Shaw, K. A., Z. L. Zhang, and N. C. MacDonald. SCREAM-I: A single mask, single crystal silicon, reactive etching process for microelectromechanical structures. Sens. Actuators A40:63–70, 1994.
Sukharev, S. I., B. Martinac, V. Y. Arshavsky, and C. Kung. Two types of mechanosensitive channels in the Escherichia coli cell envelope: Solubilization and functional reconstitution. Biophys. J.65:177–183, 1993.
Van-Essen, D. C.A tension-based theory of morphogenesis and compact wiring in the central nervous system. Nature (London)385:313–318, 1997.
Wang, N., J. P. Butler, and D. E. Ingber. Mechanotransduction across the cell surface and through the cytoskeleton. Science260:1124–1127, 1993.
Yamamoto, A., S. Mishima, N. Maruyama, and M. Sumita. A new technique for direct measurement of the shear force necessary to detach a cell from a material. Biomaterials19:871–879, 1998.
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Saif, M.T.A., Sager, C.R. & Coyer, S. Functionalized Biomicroelectromechanical Systems Sensors for Force Response Study at Local Adhesion Sites of Single Living Cells on Substrates. Annals of Biomedical Engineering 31, 950–961 (2003). https://doi.org/10.1114/1.1591189
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DOI: https://doi.org/10.1114/1.1591189