Linear Correlation between Active and Resistive Stresses Provides Information on Force Generation and Stress Transmission in Adherent Cells.

Animal cells are active, contractile objects. While bioassays address the molecular characterization of cell contractility, the mechanical characterization of the active forces in cells remains challenging. Here by confronting theoretical analysis and experiments, we calculated both the resistive and the active components of the intracellular stresses that build up following cell adhesion. We obtained a linear relationship between the divergence of the passive stress and the traction forces, which we show is the consequence of the cell adhering and applying forces on the surface only through very localized adhesion points (whose size is inferior to our best resolution, of 400 nm). This entails that there are no measurable forces outside of these active point sources, and also that the passive stresses and active stresses inside cells are proportional.

Cells on Hydrogels with Micron-Scaled Stiffness Patterns Demonstrate Local Stiffness Sensing.

Cell rigidity sensing-a basic cellular process allowing cells to adapt to mechanical cues-involves cell capabilities exerting force on the extracellular environment. In vivo, cells are exposed to multi-scaled heterogeneities in the mechanical properties of the surroundings. Here, we investigate whether cells are able to sense micron-scaled stiffness textures by measuring the forces they transmit to the extracellular matrix. To this end, we propose an efficient photochemistry of polyacrylamide hydrogels to design micron-scale stiffness patterns with kPa/µm gradients. Additionally, we propose an original protocol for the surface coating of adhesion proteins, which allows tuning the surface density from fully coupled to fully independent of the stiffness pattern. This evidences that cells pull on their surroundings by adjusting the level of stress to the micron-scaled stiffness. This conclusion was achieved through improvements in the traction force microscopy technique, e.g., adapting to substrates with a non-uniform stiffness and achieving a submicron resolution thanks to the implementation of a pyramidal optical flow algorithm. These developments provide tools for enhancing the current understanding of the contribution of stiffness alterations in many pathologies, including cancer.

Quantification of the Elastic Properties of Soft and Sticky Materials Using AFM.

Indentation Type-AFM (IT-AFM) is very useful to analyze the local rheological properties of soft or biological materials. However, analysis of the force-indentation curves is very sensitive to the way the curves are fitted: fits with elastic models such as Hertz or Sneddon's models performed on parts of the curve that indeed correspond to nonlinear elastic regimes, or that result from significant adhesive interactions of the AFM tip with the material lead to results that can be as much as twice larger than fits performed with appropriate models (nonlinear or adhesive).Here, we propose a methodology to address rigidity measurements by fitting parts of the force-indentation curves that correspond to the linear elastic response of the material, even in the presence of adhesion. The major contribution of this methodology is to set up an easy-handling criterion to mark out the linear elastic response to indentation, valid either for purely elastic and elasto-adhesive models.

AFM mapping of the elastic properties of brain tissue reveals kPa µm(-1) gradients of rigidity.

It is now well established that the mechanical environment of the cells in tissues deeply impacts cellular fate, including life cycle, differentiation and tumor progression. Designs of biomaterials already include the control of mechanical parameters, and in general, their main focus is to control the rheological properties of the biomaterials at a macroscopic scale. However, recent studies have demonstrated that cells can stress their environment below the micron scale, and therefore could possibly respond to the rheological properties of their environment at this micron scale. In this context, probing the mechanical properties of physiological cellular environments at subcellular scales is becoming critical. To this aim, we performed in vitro indentation measurements using AFM on sliced human pituitary gland tissues. A robust methodology was implemented using elasto-adhesive models, which shows that accounting for the adhesion of the probe on the tissue is critical for the reliability of the measurement. In addition to quantifying for the first time the rigidity of normal pituitary gland tissue, with a geometric mean of 9.5 kPa, our measurements demonstrated that the mechanical properties of this tissue are far from uniform at subcellular scales. Gradients of rigidity as large as 12 kPa µm(-1) were observed. This observation suggests that physiological rigidity can be highly non-uniform at the micron-scale.

Redox shuttle mechanism enhances photocatalytic H2 generation on Ni-decorated CdS nanorods.

Photocatalytic conversion of solar energy to fuels, such as hydrogen, is attracting enormous interest, driven by the promise of addressing both energy supply and storage. Colloidal semiconductor nanocrystals have been at the forefront of these efforts owing to their favourable and tunable optical and electronic properties as well as advances in their synthesis. The efficiency of the photocatalysts is often limited by the slow transfer and subsequent reactions of the photoexcited holes and the ensuing high charge recombination rates. Here we propose that employing a hydroxyl anion/radical redox couple to efficiently relay the hole from the semiconductor to the scavenger leads to a marked increase in the H2 generation rate without using expensive noble metal co-catalysts. The apparent quantum yield and the formation rate under 447 nm laser illumination exceeded 53% and 63 mmol g(-1) h(-1), respectively. The fast hole transfer confers long-term photostability on the system and opens new pathways to improve the oxidation side of full water splitting.

Resonance energy transfer improves the biological function of bacteriorhodopsin within a hybrid material built from purple membranes and semiconductor quantum dots.

Purple membrane (PM) from bacteria Halobacterium salinarum contains a photochromic protein bacteriorhodopsin (bR) arranged in a 2D hexagonal nanocrystalline lattice (Figure 1 ). Absorption of light by the protein-bound chromophore retinal results in pumping the protons through the PM creating an electrochemical gradient which is then used by the ATPases to energize the cellular processes. Energy conversion, photochromism, and photoelectrism are the inherent effects which are employed in many bR technical applications. bR, along with the other photosensitive proteins, is not able to deal with the excess energy of photons in UV and blue spectral region and utilizes less than 0.5% of the energy from the available incident solar light for its biological function. Here, we proceed with optimization of bR functions through the engineering of a "nanoconverter" of solar energy based on semiconductor quantum dots (QDs) tagged with the PM. These nanoconverters are able to harvest light from deep-UV to the visible region and to transfer this additionally collected energy to bR via Förster resonance energy transfer (FRET). We show that specific nanobio-optical and spatial coupling of QDs (donor) and bR retinal (acceptor) provide a means to achieve FRET with efficiency approaching 100%. We have finally demonstrated that the integration of QDs within PM significantly increases the efficiency of light-driven transmembrane proton pumping, which is the main bR biological function. This new QD-PM hybrid material will have numerous optoelectronic, photonic, and photovoltaic applications based on its energy conversion, photochromism, and photoelectrism properties.