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Characterization of the properties of complex biomaterials using microrheological techniques has the promise of providing fundamental insights into their biomechanical functions; however, precise interpretations of such measurements are hindered by inadequate characterization of the interactions between tracers and the networks they probe. We here show that colloid surface chemistry can profoundly affect multiple particle tracking measurements of networks of fibrin, entangled F-actin solutions, and networks of cross-linked F-actin. We present a simple protocol to render the surface of colloidal probe particles protein-resistant by grafting short amine-terminated methoxy-poly(ethylene glycol) to the surface of carboxylated microspheres. We demonstrate that these poly(ethylene glycol)-coated tracers adsorb significantly less protein than particles coated with bovine serum albumin or unmodified probe particles. We establish that varying particle surface chemistry selectively tunes the sensitivity of the particles to different physical properties of their microenvironments. Specifically, particles that are weakly bound to a heterogeneous network are sensitive to changes in network stiffness, whereas protein-resistant tracers measure changes in the viscosity of the fluid and in the network microstructure. We demonstrate experimentally that two-particle microrheology analysis significantly reduces differences arising from tracer surface chemistry, indicating that modifications of network properties near the particle do not introduce large-scale heterogeneities. Our results establish that controlling colloid-protein interactions is crucial to the successful application of multiple particle tracking techniques to reconstituted protein networks, cytoplasm, and cells.

We study the thermal motion of colloidal tracer particles in entangled actin filament (F-actin) networks, where the particle radius is comparable to the mesh size of the F-actin network. In this regime, the ensemble-averaged mean-squared displacement of the particles is proportional to tau(gamma), where 0

We measure the viscoelasticity of entangled F-actin over length scales between 1 and 100 microm using one- and two-particle microrheology, and directly identify two distinct microscopic contributions to the elasticity. Filament entanglements lead to a frequency-independent elastic modulus over an extended frequency range of 0.01-30 rad/sec; this is probed with one-particle microrheology. Longitudinal fluctuations of the filaments increase the elastic modulus between 0.1 and 30 rad/sec at length scales up to the filament persistence length; this is probed by two-particle microrheology.

We demonstrate a novel method for measuring the microrheology of soft viscoelastic media, based on cross correlating the thermal motion of pairs of embedded tracer particles. The method does not depend on the exact nature of the coupling between the tracers and the medium, and yields accurate rheological data for highly inhomogeneous materials. We demonstrate the accuracy of this method with a guar solution, for which other microscopic methods fail due to the polymer's mesoscopic inhomogeneity. Measurements in an F-actin solution suggest conventional microrheology measurements may not reflect the true bulk behavior.