Bérengère Abou

Optical tweezers reveal relationship between microstructure and nanoparticle penetration of pulmonary mucus

In this study, the mobility of nanoparticles in mucus and similar hydrogels as model systems was assessed to elucidate the link between microscopic diffusion behavior and macroscopic penetration of such gels. Differences in particle adhesion to mucus components were strongly dependent on particle coating. Particles coated with 2 kDa PEG exhibited a decreased adhesion to mucus components, whereas chitosan strongly increased the adhesion. Despite such mucoinert properties of PEG, magnetic nanoparticles of both coatings did not penetrate through native respiratory mucus, resisting high magnetic forces (even for several hours). However, model hydrogels were, indeed, penetrated by both particles in dependency of particle coating, obeying the theory of particle mobility in an external force field. Comparison of penetration data with cryogenic scanning EM images of mucus and the applied model systems suggested particularly high rigidity of the mucin scaffold and a broad pore size distribution in mucus as reasons for the observed particle immobilization. Active probing of the rigidity of mucus and model gels with optical tweezers was used in this context to confirm such properties of mucus on the microscale, thus presenting the missing link between micro- and macroscopical observations. Because of high heterogeneity in the size of the voids and pores in mucus, on small scales, particle mobility will depend on adhesive or inert properties. However, particle translocation over distances larger than a few micrometers is restricted by highly rigid structures within the mucus mesh.

Extensive collection of femtolitre pad secretion droplets in the beetle Leptinotarsa decemlineata allows nanolitre microrheology

Pads of beetles are covered with long, deformable setae, each ending in a micrometric terminal plate coated with secretory fluid. It was recently shown that the layer of the pad secretion covering the terminal plates is responsible for the generation of strong attractive forces. However, less is known about the fluid itself because it is produced in an extremely small quantity. We present here the first experimental investigation of the rheological properties of the pad secretion in the Colorado potato beetle Leptinotarsa decemlineata (Coleoptera, Chrysomelidae). Because the secretion is produced in an extremely small amount at the level of the terminal plate, we first developed a procedure based on capillary effects to collect the secretion for rheological experiments. In order to study the collected fluid (less than 1 nl) through passive microrheology, we managed to incorporate micrometric probes (melamine beads) that were initially in the form of a dry powder. Finally, the bead thermal motions were observed optically and recorded to determine the mechanical properties of the surrounding medium. We achieved this quantitative measurement with the collected volume, which is much smaller than the usual 1 µl sample volume required for this technique. Surprisingly, the beetle secretion was found to behave as a purely viscous liquid, of high viscosity (about 100 times that of water). This suggests that no specific complex fluid behaviour is needed by this adhesive system during beetle locomotion. We describe a scenario for the contact formation between the spatula at the setal tip and a smooth substrate, during the insect walk. We show that the attachment dynamics of the insect pad computed from the high measured viscosity is in good agreement with the observed insect pace. We finally discuss the consequences of the viscosity of the secretion on the insect adhesion.

Micro- and Macrorheology of Jellyfish Extracellular Matrix

Mechanical properties of the extracellular matrix (ECM) play a key role in tissue organization and morphogenesis. Rheological properties of jellyfish ECM (mesoglea) were measured in vivo at the cellular scale by passive microrheology techniques: microbeads were injected in jellyfish ECM and their Brownian motion was recorded to determine the mechanical properties of the surrounding medium. Microrheology results were compared with macrorheological measurements performed with a shear rheometer on slices of jellyfish mesoglea. We found that the ECM behaved as a viscoelastic gel at the macroscopic scale and as a much softer and heterogeneous viscoelastic structure at the microscopic scale. The fibrous architecture of the mesoglea, as observed by differential interference contrast and scanning electron microscopy, was in accord with these scale-dependent mechanical properties. Furthermore, the evolution of the mechanical properties of the ECM during aging was investigated by measuring microrheological properties at different jellyfish sizes. We measured that the ECM in adult jellyfish was locally stiffer than in juvenile ones. We argue that this stiffening is a consequence of local aggregations of fibers occurring gradually during aging of the jellyfish mesoglea and is enhanced by repetitive muscular contractions of the jellyfish.

Intracellular micro-rheology probed by micron-sized wires.

In the last decade, rapid advances have been made in the field of micro-rheology of cells and tissues. Given the complexity of living systems, there is a need for the development of new types of nano- and micron-sized probes, and in particular of probes with controlled interactions with the surrounding medium. In the present paper, we evaluate the use of micron-sized wires as potential probes of the mechanical properties of cells. The wire-based micro-rheology technique is applied to living cells such as murine fibroblasts and canine kidney epithelial cells. The mean-squared angular displacement of wires associated to their rotational dynamics is obtained as a function of the time using optical microscopy and image processing. It reveals a Brownian-like diffusive regime of the form Δψ(2)(t,L) ≈ t/L(3), where L denotes the wire length. This scaling suggests that an effective viscosity of the intracellular medium can be determined, and that in the range 1-10 μm it does not depend on the length scale over which it is measured.

3D rotational diffusion of micrometric wires using 2D video microscopy

We propose a simple way to measure the three-dimensional rotational diffusion of micrometric wires, using two-dimensional video microscopy. The out-of-plane Brownian motion of the wires in a viscous fluid is deduced from their projection on the focal plane of an optical microscope objective. An angular variable reflecting the out-of-plane motion, and satisfying a Langevin equation, is computed from the apparent wire length and its projected angular displacement. The rotational diffusion coefficient of wires between 1 and 100 μm is extracted, as well as the diameter distribution. Translational and rotational diffusion were found to be in good agreement. This is a promising way to characterize soft visco-elastic materials, and probe the dimension of anisotropic objects.

Activity statistics in a colloidal glass former: Experimental evidence for a dynamical transition

In a dense colloidal suspension at a volume fraction below the glass transition, we follow the trajectories of an assembly of tracers over a large time window. We define a local activity, which quantifies the local tendency of the system to rearrange. We determine the statistics of the time integrated activity, and we argue that it develops a low activity tail that comes together with the onset of glassy-like behavior and heterogeneous dynamics. These rare events may be interpreted as the reflection of an underlying dynamic phase transition.