Optical Tweezers Microrheology Research Articles

Nonlinear Actin Deformations Lead to Network Stiffening, Yielding, and Nonuniform Stress Propagation

We use optical tweezers microrheology and fluorescence microscopy to apply nonlinear microscale strains to entangled and cross-linked actin networks, and measure the resulting stress and actin filament deformations. We couple nonlinear stress response and relaxation to the velocities and displacements of individual fluorescent-labeled actin segments, at varying times throughout the strain and varying distances from the strain path, to determine the underlying molecular dynamics that give rise to the debated nonlinear response and stress propagation of cross-linked and entangled actin networks at the microscale. We show that initial stress stiffening arises from acceleration of strained filaments due to molecular extension along the strain, while softening and yielding is coupled to filament deceleration, halting, and recoil. We also demonstrate a surprising nonmonotonic dependence of filament deformation on cross-linker concentration. Namely, networks with no cross-links or substantial cross-links both exhibit fast initial filament velocities and reduced molecular recoil while intermediate cross-linker concentrations display reduced velocities and increased recoil. We show that these collective results are due to a balance of network elasticity and force-induced cross-linker unbinding and rebinding. We further show that cross-links dominate entanglement dynamics when the length between cross-linkers becomes smaller than the length between entanglements. In accord with recent simulations, we demonstrate that post-strain stress can be long-lived in cross-linked networks by distributing stress to a small fraction of highly strained connected filaments that span the network and sustain the load, thereby allowing the rest of the network to recoil and relax.

Time-averaged quadratic functionals of a Gaussian process

The characterization of a stochastic process from its single random realization is a challenging problem for most single-particle tracking techniques which survey an individual trajectory of a tracer in a complex or viscoelastic medium. We consider two quadratic functionals of the trajectory: the time-averaged mean-square displacement (MSD) and the time-averaged squared root mean-square displacement (SRMS). For a large class of stochastic processes governed by the generalized Langevin equation with arbitrary frictional memory kernel and harmonic potential, the exact formulas for the mean and covariance of these functionals are derived. The formula for the mean value can be directly used for fitting experimental data, e.g., in optical tweezers microrheology. The formula for the variance (and covariance) allows one to estimate the intrinsic fluctuations of measured (or simulated) time-averaged MSD or SRMS for choosing the experimental setup appropriately. We show that the time-averaged SRMS has smaller fluctuations than the time-averaged MSD, in spite of much broader applications of the latter one. The theoretical results are successfully confirmed by Monte Carlo simulations of the Langevin dynamics. We conclude that the use of the time-averaged SRMS would result in a more accurate statistical analysis of individual trajectories and more reliable interpretation of experimental data.