[HTML][HTML] Local measurements of viscoelastic parameters of adherent cell surfaces by magnetic bead microrheometry

AR Bausch, F Ziemann, AA Boulbitch, K Jacobson… - Biophysical journal, 1998 - cell.com
AR Bausch, F Ziemann, AA Boulbitch, K Jacobson, E Sackmann
Biophysical journal, 1998cell.com
A magnetic bead microrheometer has been designed which allows the generation of forces
up to 10 4 pN on 4.5μm paramagnetic beads. It is applied to measure local viscoelastic
properties of the surface of adhering fibroblasts. Creep response and relaxation curves
evoked by tangential force pulses of 500-2500pN (and∼ 1s duration) on the magnetic
beads fixed to the integrin receptors of the cell membrane are recorded by particle tracking.
Linear three-phasic creep responses consisting of an elastic deflection, a stress relaxation …
Abstract
A magnetic bead microrheometer has been designed which allows the generation of forces up to 104pN on 4.5μm paramagnetic beads. It is applied to measure local viscoelastic properties of the surface of adhering fibroblasts. Creep response and relaxation curves evoked by tangential force pulses of 500-2500pN (and ∼1s duration) on the magnetic beads fixed to the integrin receptors of the cell membrane are recorded by particle tracking. Linear three-phasic creep responses consisting of an elastic deflection, a stress relaxation, and a viscous flow are established. The viscoelastic response curves are analyzed in terms of a series arrangement of a dashpot and a Voigt body, which allows characterization of the viscoelastic behavior of the adhering cell surface in terms of three parameters: an effective elastic constant, a viscosity, and a relaxation time. The displacement field generated by the local tangential forces on the cell surface is visualized by observing the induced motion of assemblies of nonmagnetic colloidal probes fixed to the membrane. It is found that the displacement field decays rapidly with the distance from the magnetic bead. A cutoff radius of Rc∼7μm of the screened elastic field is established. Partial penetration of the shear field into the cytoplasm is established by observing the induced deflection of intracellular compartments. The cell membrane was modeled as a thin elastic plate of shear modulus μ* coupled to a viscoelastic layer, which is fixed to a solid support on the opposite side; the former accounts for the membrane/actin cortex, and the latter for the contribution of the cytoskeleton to the deformation of the cell envelope. It is characterized by the coupling constant χ characterizing the elasticity of the cytoskeleton. The coupling constant χ and the surface shear modulus μ* are obtained from the measured displacements of the magnetic and nonmagnetic beads. By analyzing the experimental data in terms of this model a surface shear modulus of μ*≈2 · 10−3Pa m to 4 · 10−3Pa m is found. By assuming an approximate plate thickness of 0.1μm one estimates an average bulk shear modulus of μ≈(2 ÷ 4) · 10−4Pa, which is in reasonable agreement with data obtained by atomic force microscopy. The viscosity of the dashpot is related to the apparent viscosity of the cytoplasm, which is obtained by assuming that the top membrane is coupled to the bottom (fixed) membrane by a viscous medium. By application of the theory of diffusion of membrane proteins in supported membranes we find a coefficient of friction of bc≈2 · 109Pa s/m corresponding to a cytoplasmic viscosity of 2 · 103Pa s.
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