Assessment of Mechanical Properties of Adherent Living Cells by Bead Micromanipulation: Comparison of Magnetic Twisting Cytometry vs Optical Tweezers

[+] Author and Article Information
Valérie M. Laurent

INSERM Unité 492, Physiopathologie et Thérapeutique Respiratoires, Faculté de Médecine et Faculté des Sciences et Technologie, Université Paris XII, 8, rue du Général Sarrail, 94010 CRÉTEIL cedex France CNRS, UMR-7057 associé aux Universités Paris VI et Paris VII, Laboratoire de Biorhéologie et d’Hydrodynamique Physicochimique, et Fédération de Recherche Matière et Systèmes Complexes, 2 place Jussieu, 75251 PARIS cedex 5, France

Sylvie Hénon, Martial Balland, François Gallet

CNRS, ESA-7057 associé aux Universités Paris VI et Paris VII, Laboratoire de Biorhéologie et d’Hydrodynamique Physicochimique, 2 place Jussieu, 75251 PARIS cedex 5, France

Emmanuelle Planus, Redouane Fodil

INSERM Unité 492, Physiopathologie et at Thérapeutique Respiratoires, Faculté de Médecine et Faculté des Sciences et Technologie, Université Paris XII, 8, rue du Général Sarrail, 94010 CRÉTEIL cedex France

Daniel Isabey

INSERM Unité 492, Physiopathologie et Thérapeutique Respiratoires, Faculté de Médecine et Faculté des Sciences et Technologie, Université Paris XII, 8, rue du Général Sarrail, 94010 CRÉTEIL cedex France

J Biomech Eng 124(4), 408-421 (Jul 30, 2002) (14 pages) doi:10.1115/1.1485285 History: Received November 29, 2000; Revised November 05, 2001; Online July 30, 2002
Copyright © 2002 by ASME
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Dai,  J., and Sheetz,  M. P., 1995, “Regulation of Endocytosis, exocytosis, and shape by Membrane Tension,” Cold Spring Harbor Symp. Quant. Biol., 60, pp. 567–571.
Davies,  P. F., and Tripathi,  S. C., 1993, “Mechanical Stress Mechanisms and the Cell, an Endothelial Paradigm,” Circ. Res., 72, pp. 239–245.
Heidemann,  S. R., and Buxbaum,  R. E., 1994, “Mechanical Tension as a Regulator of Axonal Development,” Neurotoxicology, 15, pp. 95–107.
Ingber,  D. E., Prusty,  D., Sun,  Z., Betensky,  H., and Wang,  N., 1995, “Cell Shape, Cytoskeletal Mechanics, and Cell Cycle Control in Angiogenesis,” J. Biomech., 28, pp. 1471–1484.
Schmidt,  C. E., Dai,  J., Lauffenburger,  D. A., Sheetz,  M. P., and Horwitz,  A. F., 1995, “Integrin-cytoskeletal Interactions in Neuronal Growth Cones,” J. Neurosci., 15, pp. 3400–3407.
Stossel,  T. P., 1993, “On the Crawling of Animal Cells,” Science, 260, pp. 1086–1094.
Wang,  N., and Ingber,  D. E., 1994, “Control of Cytoskeletal Mechanics by Extracellular Matrix, Cell Shape and Mechanical Tension,” Biophys. J., 66, pp. 1–9.
Wang,  N., Butler,  J. P., and Ingber,  D. E., 1993, “Mechanotransduction Across the Cell Surface and Through the Cytoskeleton [see comments],” Science, 260, pp. 1124–1127.
Choquet,  D., Felsenfeld,  D. P., and Sheetz,  M. P., 1997, “Extracellular Matrix Rigidity Causes Strengthening of Integrin-Cytoskeleton Linkages,” Cell, 88, pp. 39–48.
Heidemann,  S. R., 1993, “A New Twist on Integrins and the Cytoskeleton,” Science, 260, pp. 1080–1081.
Ingber,  D. E., Dike,  L., Hansen,  L., Karp,  S., Liley,  H., Maniotis,  A., McNamee,  H., Mooney,  D., Plopper,  G., Sims,  J., and Wang,  N., 1994, “Cellular Tensegrity: Exploring How Mechanical Changes in the Cytoskeleton Regulate Cell Growth, Migration, and Tissue Pattern During Morphogenesis,” Int. Rev. Cytol. 150, pp. 173–224.
Janmey,  P. A., 1998, “The Cytoskeleton and Cell Signaling: Component Localization and Mechanical Coupling,” Physiol. Rev., 78, pp. 763–781.
Planus,  E., Galiacy,  S., Matthay,  M., Laurent,  V., Gavrilovic,  J., Murphy,  G., Clérici,  C., Isabey,  D., Lafuma,  C., and d’Ortho,  M. P., 1999, “Role of Collagenase in Mediating In Vitro Alveolar Epithelial Wound Repair,” J. Cell. Sci., 112(Pt. 2), pp. 243–252.
Potard,  U. S., Butler,  J. P., and Wang,  N., 1997, “Cytoskeletal Mechanics in Confluent Epithelial Cells Probed through Integrins and E-cadherins,” Am. J. Physiol., 272, pp. C1654–1663.
Hubmayr,  R. D., Shore,  S. A., Fredberg,  J. J., Planus,  E., and Wang,  N., 1996, “Pharmacological Activation Changes Stiffness of Cultured Human Airway Smooth Muscle Cells,” Am. J. Physiol., 271, pp. C1660–C1668.
Sato,  M., Theret,  D. P., Wheeler,  L. T., Ohshima,  N., and Nerem,  R. M., 1990, “Application of the Micropipette Technique to the Measurement of Cultured Porcine Aortic Endothelial Cell Viscoelastic Properties,” J. Biomech. Eng., 112, pp. 263–268.
Sato,  M., Ohshima,  N., and Nerem,  R. M., 1996, “Viscoelastic Properties of Cultured Porcine Aortic Endothelial Cells Exposed to Shear Stress,” J. Biomech., 29, pp. 461–467.
Petersen,  N. O., McConnaughey,  W. B., and Elson,  E. L., 1982, “Dependence of Locally Measured Cellular Deformability on Position on the Cell, Temperature, and Cytochalasin B,” Proc. Natl. Acad. Sci. U.S.A., 79, pp. 5327–5331.
Thoumine,  O., Ott,  A., Cardoso,  O., and Meister,  J. J., 1999, “Microplates: A New Tool for Manipulation and Mechanical Perturbation of Individual Cells,” J. Biochem. Biophys. Methods, 39, pp. 47–62.
Bausch,  A. R., Möller,  W., and Sackmann,  E., 1999, “Measurement of Local Viscoelasticity and Forces in Living Cells by Magnetic Tweezers,” Biophys. J., 76, pp. 573–579.
Shroff,  S. G., Saner,  D. R., and Lal,  R., 1995, “Dynamic Micromechanical Properties of Cultured Rat Atrial Myocytes Measured by Atomic Force Microscopy,” Am. J. Physiol., 269, pp. C286–292.
Stamenovic,  D., and Coughlin,  M. F., 2000, “A Quantitative Model of Cellular Elasticity Based on Tensegrity,” J. Biomech. Eng., 122, pp. 39–43.
Wendling,  S., Oddou,  C., and Isabey,  D., 1999, “Stiffening Response of a Cellular Tensegrity Model,” J. Theor. Biol., 196, pp. 309–325.
Svoboda,  K., and Block,  S. M., 1994, “Biological Applications of Optical Forces,” Annu. Rev. Biophys. Biomol. Struct., 23, pp. 247–285.
Hénon,  S., Lenormand,  G., Richert,  A., and Gallet,  F., 1999, “A New Determination of the Shear Modulus of the Human Erythrocyte Membrane Using Optical Tweezers,” Biophys. J., 76, pp. 1145–1151.
Lenormand,  G., Hénon,  S., and Gallet,  F., 1999, “Measurement of the Elastic Coefficients of an Isolated Red Blood Cell Membrane Skeleton with Optical Tweezers,” Arch. Physiol. Biochem., 107, pp. 69.
Wang,  N., and Ingber,  D. E., 1995, “Probing Transmembrane Mechanical Coupling and Cytomechanics Using Magnetic Twisting Cytometry,” Biochem. Cell Biol., 73, pp. 327–335.
Wendling,  S., Planus,  E., Laurent,  V., Barbe,  L., Mary,  A., Oddou,  C., and Isabey,  D., 2000, “Role of Cellular Tone and Microenvironmental on Cytoskeleton Stiffness Assessed by Tensegrity Model,” Eur. Phys. J.: Appl. Phys., 9, pp. 51–62.
Yamada,  S., Wirtz,  D., and Kuo,  S. C., 2000, “Mechanics of Living Cells Measured by Laser Tracking Microrheology,” Biophys. J., 78, pp. 1736–1747.
Foner,  S., and Macniff Jr,  E. J., 1968, The Review of Scientific Instrument., 39, pp. 171–181.
Sheetz,  M. P., 1998, “Laser Tweezers in Cell Biology, Introduction,” Methods Cell Biol., 55, pp. xi–xii.
Simmons,  R. M., Finer,  J. T., Chu,  S., and Spudich,  J. A., 1996, “Quantitative Measurements of Force and Displacement Using an Optical Trap,” Biophys. J., 70, pp. 1813–1822.
Wang,  N., 1998, “Mechanical Interactions Among Cytoskeletal Filaments,” Hypertension, 32, pp. 162–165.
Stamenovic,  D., and Coughlin,  M. F., 1999, “The Role of Prestress and Architecture of the Cytoskeleton and Deformability of Cytoskeleton Filaments in Mechanics of Adherent Cells: A Quantitative Analysis,” J. Theor. Biol., 201, pp. 63–74.
Ziemann,  F., Rädler,  J., and Sackmann,  E., 1994, “Local Measurements of Viscoelastic Moduli of Entangled Actin Networks Using an Oscillating Magnetic Bead Micro-rheometer,” Biophys. J., 66, pp. 2210–2216.
Bausch,  A. R., Ziemann,  F., Boulbitch,  A. A., Jacobson,  K., and Sackmann,  E., 1998, “Local Measurements of Viscoelastic Parameters of Adherent Cell Surfaces by Magnetic Bead Microrheometry,” Biophys. J., 75, pp. 2038–2049.
Maksym,  G. N., Fabry,  B., Butler,  J. P., Navajas,  D., Tschumperlin,  D. J., Laporte,  J. D., and Fredberg,  J. J., 2000, “Mechanical Properties of Cultured Human Airway Smooth Muscle Cells from 0.05 to 0.4 Hz,” J. Appl. Physiol., 89, pp. 1619–1632.
Rotsch,  C., Braet,  F., Wisse,  E., and Radmacher,  M., 1997, “AFM Imaging and Elasticity Measurements on Living Rat Liver Macrophages,” Cell Biol. Int., 21, pp. 685–696.
Goldmann,  W. H., Galneder,  R., Ludwig,  M., Lu,  W., Adamson,  E. D., Wang,  N., and Ezzell,  R. M., 1998, “Differences in Elasticity of Vinculin-deficient F9 Cells Measured by Magnetometry and Atomic Force Microscopy,” Exp. Cell Res., 239, pp. 235–242.
Maniotis,  A. J., Chen,  C. S., and Ingber,  D. E., 1997, “Demonstration of Mechanical Connections Between Integrins, Cytoskeletal Filaments, and Nucleoplasm that Stabilize Nuclear Structure,” Proc. Natl. Acad. Sci. U.S.A., 94, pp. 849–854.
Fabry,  B., Maksym,  G. N., Hubmayr,  R. D., Butler,  J. P., and Fredberg,  J. J., 1999, “Implications of Heterogeneous Bead Behavior on Cell Mechanical Properties Measured with Magnetic Twisting Cytometry,” J. Magn. Magn. Mater., 194, pp. 120–125.
Richelme,  F., Benoliel,  A. M., and Bongrand,  P., 2000, “Dynamic Study of Cell Mechanical and Structural Responses to Rapid Changes of Calcium Level,” Cell Motil. Cytoskeleton, 45, pp. 93–105.
Wang,  J. S., Pavlotsky,  N., Tauber,  A. I., and Zaner,  K. S., 1993, “Assembly Dynamics of Actin in Adherent Human Neutrophils,” Cell Motil. Cytoskeleton, 26, pp. 340–348.
Landau, L., and Lifchitz, E., 1959, Theory of Elasticity, Pergamon Press, London.
Love, A. E. H., 1944, A Treatise on the Mathematical Theory of Elasticity (4th edition), Dover Publications, New York.
Landau, L., and Lifchitz, E., 1971, Fluid Mechanics, Bergamon Press, London.


Grahic Jump Location
Plot of the position x (in μm) of the bead center versus applied force F (in pN), characterizing optical tweezers experiments. The accuracy of the measurements is about ±0.1 μm for x and ±5% for F. The x-F relationship is mostly linear in contrast with magnetic twisting cytometry experiments.
Grahic Jump Location
The equivalent Young modulus E (in Pa) for 24 different epithelial alveolar cells measured with optical tweezers. The measured values of E lie in the range 29–258 Pa, with an average value at 125 Pa.
Grahic Jump Location
Plot of the relaxation of the position x (in μm) of a bead versus time (in seconds) when the force applied by optical tweezers is switched off. The best mono-exponential fit is also shown and corresponds to a relaxation time constant: τ=3.5 s close to the mean value of 3.1±2.0 s found over 21 similar relaxation curve experiments.
Grahic Jump Location
Experimental comparison of the data obtained with magnetic twisting cytometry and optical tweezers: the rotation component θ° (bead deviation angle in degrees) is plotted against the external torque C (in pN×μm) applied at equilibrium, for each micromanipulation technique. For optical tweezers, the results obtained for three different measurements (upper (□); intermediate (×); lower values (○) of equivalent Young modulus) are shown and exhibit three linear θ°-C relationships, e.g., from bottom to top: E=230 Pa (close to the highest value of E measured with optical tweezers), E=140 Pa (close to the average value), E=30 Pa (close to the lowest measured value). The estimated accuracy is about ±1° for the deviation angle θ°. Magnetic twisting cytometry generally generates larger bead deviations θ° (values in degrees are mean ±SE and correspond to dark circles •), associated with a larger range of applied external torque C. Accordingly, the corresponding θ°-C relationship appears to be fairly nonlinear with twisting magnetic cytometry (see Discussion).
Grahic Jump Location
Various geometric configurations for a bead bound to a cell considered to be a homogeneous elastic medium: (a) The bead is fully immersed in an infinite three-dimensional medium; (b) Half the bead is immersed in a semi-infinite medium; (c) The bead is in contact with the flat surface of a semi-infinite medium via a small circular area. The degree of bead immersion is measured by the half-angle α generating the cone limiting the contact area.
Grahic Jump Location
Spatial reconstruction of the bead/F-actin structure using confocal microscopic images obtained after staining with fluorescent phallotoxin. The bead/F-actin structure is represented here for two beads (a) and (b) corresponding to two vertical sections in the same epithelial cell and in conditions representative of magnetic twisting cytometry measurements. Contact points between the bead and the F-actin structure allowed accurate measurements of the half immersion angle α, e.g., α=78.2° and α=66.8° for these two beads (see text for explanations).
Grahic Jump Location
Bead immersion angle (α) in the cytoplasm, measured in (a) from the vertical sections of 3D-reconstructed images of bead/F-actin structure for 25 beads representative of magnetic twisting experiments, in (b) from brightfield microscopic images taken in the actual conditions of the laser trapping, namely for 24 beads. Note the similarities between the distribution angle α measured with two different methods throughout an equivalent number of beads. The mean value of α measured in each technique, i.e., 67° in magnetic twisting and 62° in laser trapping experiments, are fairly compatible.
Grahic Jump Location
Plot of the bead deviation angle θ° (in degrees) versus the applied magnetic torque C (in pN×μm). The θ°-C relationship is nonlinear, as previously reported in magnetic twisting cytometry experiments. Values are mean ±SD.
Grahic Jump Location
Plot of the equivalent Young modulus E (in Pascal) for the four values of applied magnetic torque C (in pN×μm), calculated using Eq. (13) with α=αm=67° for a typical magnetic twisting cytometry experiment. The E-values lie in the range 34–58 Pa, and increase as magnetic torque increases. Values are mean ±SD.
Grahic Jump Location
Plot of the bead rotation angle θt (in degrees) versus time (in seconds) in the relaxation period, i.e., when the magnetic torque is released. Experimental data correspond to the gray points. The best mono-exponential fit is also shown (dark curve) and corresponds to a relaxation time constant τ=1.9 s close to the mean value of 1.3±0.8 s found over 15 similar relaxation curve experiments. Note that the final equilibrium state (θ≈22°) differs from the initial state (θ=0°).
Grahic Jump Location
Images recorded during an optical tweezers experiment. (a): a silica microbead bound at rest to an epithelial alveolar cell; (b): a force F≈240 pN is applied to the bead by the optical tweezers in a direction parallel to the cell membrane; (c): superimposition of (a) and (b): bead rotation is θ≈5° and total translation of the bead center is x≈0.8 μm; scale given by the bead diameter (5 μm).




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