Nanorheology and Nanoindentation Revealed a Softening and an Increased Viscous Fluidity of Adherent Mammalian Cells upon Increasing the Frequency.

The nanomechanical response of a cell depends on the frequency at which the cell is probed. The components of the cell that contribute to this property and their interplay are not well understood. Here, two force microscopy methods are integrated to characterize the frequency and/or the velocity-dependent properties of living cells. It is shown on HeLa and fibroblasts, that cells soften and fluidize upon increasing the frequency or the velocity of the deformation. This property was independent of the type and values (25 or 1000 nm) of the deformation. At low frequencies (2-10 Hz) or velocities (1-10 µm s-1 ), the response is dominated by the mechanical properties of the cell surface. At higher frequencies (>10 Hz) or velocities (>10 µm s-1 ), the response is dominated by the hydrodynamic drag of the cytosol. Softening and fluidization does not seem to involve any structural remodeling. It reflects a redistribution of the applied stress between the solid and liquid-like elements of the cell as the frequency or the velocity is changed. The data indicates that the quasistatic mechanical properties of a cell featuring a cytoskeleton pathology might be mimicked by the response of a non-pathological cell which is probed at a high frequency.

Fast and high-resolution mapping of van der Waals forces of 2D materials interfaces with bimodal AFM.

High-spatial resolution mapping of van der Waals forces is relevant in several fields ranging from nanotechnology to colloidal science. The emergence of two-dimensional heterostructures assembled by van der Waals interactions has enhanced the interest of those measurements. Several AFM methods have been developed to measure the adhesion force between an AFM probe and the material of interest. However, a reliable and high-resolution method to measure the Hamaker constant remains elusive. We demonstrate that an atomic force microscope operated in a bimodal configuration enables fast, quantitative, and high-resolution mapping of the Hamaker constant of interfaces. The method is applied to map the Hamaker constant of monolayer, bilayer and multilayer MoS2 surfaces. Those interfaces are characterized with Hamaker constant and spatial resolutions of, respectively, 0.1 eV and 50 nm.

Insights and guidelines to interpret forces and deformations at the nanoscale by using a tapping mode AFM simulator: dForce 2.0.

Amplitude modulation (tapping mode) AFM is the most versatile AFM mode for imaging surfaces at the nanoscale in air and liquid environments. However, it remains challenging to estimate the forces and deformations exerted by the tip. We introduce a new simulator environment to predict the values of the observables in tapping mode AFM experiments. The relevant feature of dForce 2.0 is the incorporation of contact mechanics models aimed to describe the properties of ultrathin samples. These models were essential to determine the forces applied on samples such as proteins, self-assembled monolayers, lipid bilayers, and few-layered materials. The simulator incorporates two types of long-range magnetic forces. The simulator is written in an open-source code (Python) and it can be run from a personal computer.

Accurate Wide-Modulus-Range Nanomechanical Mapping of Ultrathin Interfaces with Bimodal Atomic Force Microscopy.

The nanoscale determination of the mechanical properties of interfaces is of paramount relevance in materials science and cell biology. Bimodal atomic force microscopy (AFM) is arguably the most advanced nanoscale method for mapping the elastic modulus of interfaces. Simulations, theory, and experiments have validated bimodal AFM measurements on thick samples (from micrometer to millimeter). However, the bottom-effect artifact, this is, the influence of the rigid support on the determination of the Young's modulus, questions its accuracy for ultrathin materials and interfaces (1-15 nm). Here we develop a bottom-effect correction method that yields the intrinsic Young's modulus value of a material independent of its thickness. Experiments and numerical simulations validate the accuracy of the method for a wide range of materials (1 MPa to 100 GPa). Otherwise, the Young's modulus of an ultrathin material might be overestimated by a 10-fold factor.

Quantitative mapping of magnetic properties at the nanoscale with bimodal AFM.

We demonstrate that a force microscope operated in a bimodal configuration enables the mapping of magnetic interactions with high quantitative accuracy and high-spatial resolution (∼30 nm). Bimodal AFM operation doubles the number of observables with respect to conventional magnetic force microscopy methods which enables to determine quantitatively in a single processing step several magnetic properties. The theory of bimodal AFM provides analytical expressions for different magnetic force models, in particular those characterized by power-law and exponential distance dependences. Bimodal AFM provides a self-evaluation protocol to test the accuracy of the measurements. The agreement obtained between the experiments and theory for two different magnetic samples support the application of bimodal AFM to map quantitatively long-range magnetic interactions.

High-Speed Nanomechanical Mapping of the Early Stages of Collagen Growth by Bimodal Force Microscopy.

High-speed atomic force microscopy (AFM) enabled the imaging of protein interactions with millisecond time resolutions (10 fps). However, the acquisition of nanomechanical maps of proteins is about 100 times slower. Here, we developed a high-speed bimodal AFM that provided high-spatial resolution maps of the elastic modulus, the loss tangent, and the topography at imaging rates of 5 fps. The microscope was applied to identify the initial stages of the self-assembly of the collagen structures. By following the changes in the physical properties, we identified four stages, nucleation and growth of collagen precursors, formation of tropocollagen molecules, assembly of tropocollagens into microfibrils, and alignment of microfibrils to generate microribbons. Some emerging collagen structures never matured, and after an existence of several seconds, they disappeared into the solution. The elastic modulus of a microfibril (∼4 MPa) implied very small stiffness (∼3 × 10-6 N/m). Those values amplified the amplitude of the collagen thermal fluctuations on the mica plane, which facilitated microribbon build-up.

Atrial location optimization by electrical measures for Electrocardiographic Imaging.

BACKGROUND: The Electrocardiographic Imaging (ECGI) technique, used to non-invasively reconstruct the epicardial electrical activity, requires an accurate model of the atria and torso anatomy. Here we evaluate a new automatic methodology able to locate the atrial anatomy within the torso based on an intrinsic electrical parameter of the ECGI solution. METHODS: In 28 realistic simulations of the atrial electrical activity, we randomly displaced the atrial anatomy for ±2.5 cm and ±30° on each axis. An automatic optimization method based on the L-curve curvature was used to estimate the original position using exclusively non-invasive data. RESULTS: The automatic optimization algorithm located the atrial anatomy with a deviation of 0.5 ± 0.5 cm in position and 16.0 ± 10.7° in orientation. With these approximate locations, the obtained electrophysiological maps reduced the average error in atrial rate measures from 1.1 ± 1.1 Hz to 0.5 ± 1.0 Hz and in the phase singularity position from 7.2 ± 4.0 cm to 1.6 ± 1.7 cm (p < 0.01). CONCLUSIONS: This proposed automatic optimization may help to solve spatial inaccuracies provoked by cardiac motion or respiration, as well as to use ECGI on torso and atrial anatomies from different medical image systems.

Combined Magnetoliposome Formation and Drug Loading in One Step for Efficient Alternating Current-Magnetic Field Remote-Controlled Drug Release.

We have developed a reproducible and facile one step strategy for the synthesis of doxorubicin loaded magnetoliposomes by using a thin-layer evaporation method. Liposomes of around 200 nm were made of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and iron oxide nanoparticles (NPs) with negative, positive, and hydrophobic surfaces that were incorporated outside, inside, or between the lipid bilayers, respectively. To characterize how NPs are incorporated in liposomes, advanced cryoTEM and atomic force microscope (AFM) techniques have been used. It was observed that only when the NPs are attached outside the liposomes, the membrane integrity is preserved (lipid melt transition shifts to 38.7 °C with high enthalpy 34.8 J/g) avoiding the leakage of the encapsulated drug while having good colloidal properties and the best heating efficiency under an alternating magnetic field (AMF). These magnetoliposomes were tested with two cancer cell lines, MDA-MB-231 and HeLa cells. First, 100% of cellular uptake was achieved with a high cell survival (above 80%), which is preserved (83%) for doxorubicin-loaded magnetoliposomes. Then, we demonstrate that doxorubicin release can be triggered by remote control, using a noninvasive external AMF for 1 h, leading to a cell survival reduction of 20%. Magnetic field conditions of 202 kHz and 30 mT seem to be enough to produce an effective heating to avoid drug degradation. In conclusion, these drug-loaded magnetoliposomes prepared in one step could be used for drug release on demand at a specific time and place, efficiently using an external AMF to reduce or even eliminate side effects.

Phasa: todo sobre la arcilla comestible / Phasa: everything about edible clay

La phasa (conocida así entre los pobladores aymaras) es un tipo de arcilla comestible que se encuentra en los suelos, empleada como medicina tradicional en la mayoría de los continentes, principalmente para aliviar o tratar patologías gastrointestinales e infecciones cutáneas. Su consumo se remonta muchos siglos atrás; existe evidencia del uso de este elemento en las Placas de arcilla de Nippur en Mesopotamia (2500 años a. C.). Develar las propiedades bio-físico-químicas de las arcillas comestibles está permitiendo a la ciencia explicar las propiedades terapéuticas que posee y así confirmar los grandes avances en la medicina alternativa que tuvieron nuestros antepasados. El uso de la phasa no se limita a la medicina, sino también a muchas otras ramas de la ciencia. En los últimos 10 años se ha experimentado en la agricultura, veterinaria, incluso en el cuidado del medio ambiente dando resultados muy gratificantes y prometedores.

Phasa, an Aymara's language word, is a type of edible clay found in soils; which is used in traditional medicine worldwide mainly to relieve or treat gastrointestinal pathologies and skin infections. Its consumption dates back many centuries, even millennia ago; there is evidence of the use of this element in the clay plates of Nippur in Mesopotamia (2500 years BC). Revealing the bio-physical-chemical properties of edible clay has allowed science to explain their healing and therapeutic properties and confirms the great advances our ancestors got. The use of phasa is not limited to medicine, it is also used in many other sciences; in the last 10 years it has been used in agriculture and veterinary medicine, getting gratifying results.

Fast and high-resolution mapping of elastic properties of biomolecules and polymers with bimodal AFM.

Fast, high-resolution mapping of heterogeneous interfaces with a wide elastic modulus range is a major goal of atomic force microscopy (AFM). This goal becomes more challenging when the nanomechanical mapping involves biomolecules in their native environment. Over the years, several AFM-based methods have been developed to address this goal. However, none of these methods combine sub-nanometer spatial resolution, quantitative accuracy, fast data acquisition speed, wide elastic modulus range and operation in physiological solutions. Here, we present detailed procedures for generating high-resolution maps of the elastic properties of biomolecules and polymers using bimodal AFM. This requires the simultaneous excitation of the first two eigenmodes of the cantilever. An amplitude modulation (AM) feedback acting on the first mode controls the tip-sample distance, and a frequency modulation (FM) feedback acts on the second mode. The method is fast because the elastic modulus, deformation and topography images are obtained simultaneously. The method is efficient because only a single data point per pixel is needed to generate the aforementioned images. The main stages of the bimodal imaging are sample preparation, calibration of the instrument, tuning of the microscope and generation of the nanomechanical maps. In addition, with knowledge of the deformation, bimodal AFM enables reconstruction of the true topography of the surface. It takes ~9 h to complete the whole procedure.