The cell biologist's guide to super-resolution microscopy

Fluorescence microscopy has become a ubiquitous method to observe the location of specific molecular components within cells. However, the resolution of light microscopy is limited by the laws of diffraction to a few hundred nanometers, blurring most cellular details. Over the last two decades, several techniques – grouped under the ‘super-resolution microscopy’ moniker – have been designed to bypass this limitation, revealing the cellular organization down to the nanoscale. The number and variety of these techniques have steadily increased, to the point that it has become difficult for cell biologists and seasoned microscopists alike to identify the specific technique best suited to their needs. Available techniques include image processing strategies that generate super-resolved images, optical imaging schemes that overcome the diffraction limit and sample manipulations that expand the size of the biological sample. In this Cell Science at a Glance article and the accompanying poster, we provide key pointers to help users navigate through the various super-resolution methods by briefly summarizing the principles behind each technique, highlighting both critical strengths and weaknesses, as well as providing example images

Minimising pain in farm animals: the 3S approach - ‘Suppress, Substitute, Soothe'

Recently, the French National Institute for Agricultural Research appointed an expert committee to review the issue of pain in food-producing farm animals. To minimise pain, the authors developed a ‘3S' approach accounting for ‘Suppress, Substitute and Soothe' by analogy with the ‘3Rs' approach of ‘Reduction, Refinement and Replacement' applied in the context of animal experimentation. Thus, when addressing the matter of pain, the following steps and solutions could be assessed, in the light of their feasibility (technical constraints, logistics and regulations), acceptability (societal and financial aspects) and availability. The first solution is to suppress any source of pain that brings no obvious advantage to the animals or the producers, as well as sources of pain for which potential benefits are largely exceeded by the negative effects. For instance, tail docking of cattle has recently been eliminated. Genetic selection on the basis of resistance criteria (as e.g. for lameness in cattle and poultry) or reduction of undesirable traits (e.g. boar taint in pigs) may also reduce painful conditions or procedures. The second solution is to substitute a technique causing pain by another less-painful method. For example, if dehorning cattle is unavoidable, it is preferable to perform it at a very young age, cauterising the horn bud. Animal management and constraint systems should be designed to reduce the risk for injury and bruising. Lastly, in situations where pain is known to be present, because of animal management procedures such as dehorning or castration, or because of pathology, for example lameness, systemic or local pharmacological treatments should be used to soothe pain. These treatments should take into account the duration of pain, which, in the case of some management procedures or diseases, may persist for longer periods. The administration of pain medication may require the intervention of veterinarians, but exemptions exist where breeders are allowed to use local anaesthesia (e.g. castration and dehorning in Switzerland). Extension of such exemptions, national or European legislation on pain management, or the introduction of animal welfare codes by retailers into their meat products may help further developments. In addition, veterinarians and farmers should be given the necessary tools and information to take into account animal pain in their management decision

Análisis de la nitración fisiológica de proteínas en órganos de plantas de guisante (Pisum sativum L) durante su desarrollo y senescencia

Comunicaciones a congreso

Nonlinear Elasticity in Biological Gels

Unlike most synthetic materials, biological materials often stiffen as they are deformed. This nonlinear elastic response, critical for the physiological function of some tissues, has been documented since at least the 19th century, but the molecular structure and the design principles responsible for it are unknown. Current models for this response require geometrically complex ordered structures unique to each material. In this Article we show that a much simpler molecular theory accounts for strain stiffening in a wide range of molecularly distinct biopolymer gels formed from purified cytoskeletal and extracellular proteins. This theory shows that systems of semi-flexible chains such as filamentous proteins arranged in an open crosslinked meshwork invariably stiffen at low strains without the need for a specific architecture or multiple elements with different intrinsic stiffnesses.Comment: 23 pages, 5 figures, submitted to Natur

Elongation and fluctuations of semi-flexible polymers in a nematic solvent

We directly visualize single polymers with persistence lengths ranging from $\ell_p=0.05$ to 16 $\mu$m, dissolved in the nematic phase of rod-like {\it fd} virus. Polymers with sufficiently large persistence length undergo a coil-rod transition at the isotropic-nematic transition of the background solvent. We quantitatively analyze the transverse fluctuations of semi-flexible polymers and show that at long wavelengths they are driven by the fluctuating nematic background. We extract both the Odijk deflection length and the elastic constant of the background nematic phase from the data.Comment: 4 pages, 4 figures, submitted to PR

Mechanics of fragmentation of crocodile skin and other thin films

Fragmentation of thin layers of materials is mediated by a network of cracks on its surface. It is commonly seen in dehydrated paintings or asphalt pavements and even in graphene or other two-dimensional materials, but is also observed in the characteristic polygonal pattern on a crocodile’s head. Here, we build a simple mechanical model of a thin film and investigate the generation and development of fragmentation patterns as the material is exposed to various modes of deformation. We find that the characteristic size of fragmentation, defined by the mean diameter of polygons, is strictly governed by mechanical properties of the film material. Our result demonstrates that skin fragmentation on the head of crocodiles is dominated by that it features a small ratio between the fracture energy and Young’s modulus, and the patterns agree well with experimental observations. Understanding this mechanics-driven process could be applied to improve the lifetime and reliability of thin film coatings by mimicking crocodile skin

Frequency dependent dielectric and mechanical behaviour of elastomers for actuator applications

The low frequency mechanical and dielectric behavior of three different elastomers has been investigated by dynamic mechanical analysis and dielectric spectroscopy, with the aim of accounting for the frequency dependence of the characteristics of the corresponding dielectric elastomer actuators. Satisfactory agreement was obtained between the dynamic response of the actuators and a simple model based on the experimental data for the elastomers, assuming that the relatively large prestrains employed in the actuators to have little influence on the frequency dependence of their effective moduli. It was thus demonstrated that the frequency dependence of the actuator strain is dominated by that of the mechanical response of the elastomer, and that the frequency dependence of the dielectric properties has a relatively minor influence on the actuator performance

Cohesion and adhesion of nanoporous TiO2 coatings on titanium wires for photovoltaic applications

The working electrode of a dye-sensitized photovoltaic fiber is constituted of a porous TiO2 coated titanium wire. The cohesion and adhesion of such a brittle oxide coating on the ductile metal substrate are identified as crucial factors in maintaining photovoltaic efficiency during textile manufacture and weaving operations. The influence of coating thickness on these factors has been investigated in the present work. The tensile mechanical characterization with in situ microscopic observations shows that two damaging processes are involved. For the smaller thickness, loss of adherence appears to be at the interface and inside the coating bulk. Cracks become visible in a random distribution in size and density and do not cross the entire coating circumference. Large patches of coating are still anchored on the wire. For the larger thickness no cohesive rupture in the coating bulk has been observed. The loss of adherence appears at the interface closed to the cracks and grows rapidly as the strain increases. Numerical investigations based on the finite element method permit to analyze the distribution and the combination of radial interfacial stress and circumferential coating stress and their influence on the observed damage