53 research outputs found
Nanomechanical and topographical imaging of living cells by Atomic Force Microscopy with colloidal probes
Atomic Force Microscopy (AFM) has a great potential as a tool to characterize
mechanical and morphological properties of living cells; these properties have
been shown to correlate with cells' fate and patho-physiological state in view
of the development of novel early-diagnostic strategies. Although several
reports have described experimental and technical approaches for the
characterization of cell elasticity by means of AFM, a robust and commonly
accepted methodology is still lacking. Here we show that micrometric spherical
probes (also known as colloidal probes) are well suited for performing a
combined topographic and mechanical analysis of living cells, with spatial
resolution suitable for a complete and accurate mapping of cell morphological
and elastic properties, and superior reliability and accuracy in the mechanical
measurements with respect to conventional and widely used sharp AFM tips. We
address a number of issues concerning the nanomechanical analysis, including
the applicability of contact mechanical models and the impact of a constrained
contact geometry on the measured elastic modulus (the finite-thickness effect).
We have tested our protocol by imaging living PC12 and MDA-MB-231 cells, in
order to demonstrate the importance of the correction of the finite-thickness
effect and the change in cell elasticity induced by the action of a
cytoskeleton-targeting drug.Comment: 51 pages, 12 figures, 3 table
Ultrasound tissue characterization detectspreclinical myocardial structural changes inchildren affected by Duchenne muscular dystrophy
AbstractObjectivesOur goal was to identify early changes in myocardial physical properties in children with Duchenne muscular dystrophy (DMDch).BackgroundDuchenne muscular dystrophy (DMD) is caused by the absence of dystrophin, which triggers complex molecular and biological events in skeletal and cardiac muscle tissues. Although about 30% of patients display overt signs of cardiomyopathy in the late stage of the disease, it is unknown whether changes in myocardial physical properties can be detected in the early (preclinical) stages of the disease.MethodsWe performed an ultrasonic tissue characterization (UTC) analysis of myocardium in DMDch with normal systolic myocardial function and no signs of cardiomyopathy. Both the cyclic variation of integrated backscatter (cvIBS) and the calibrated integrated backscatter (cIBS) were assessed in 8 myocardial regions of 20 DMDch, age 7 ± 2 years (range 4 to 10 years), and in 20 age-matched healthy controls.ResultsWe found large differences in the UTC data between DMDch and controls; the mean value of cvIBS was 4.4 ± 1.5 dB versus 8.8 ± 0.8 dB, whereas the mean value of cIBS was 36.4 ± 7.1 dB versus 26.9 ± 2.0 dB (p < 10−6for both). In DMDch, all eight sampled segments showed cIBS mean values to be significantly higher and cvIBS mean values to be significantly lower than those in the controls. Finally, interindividual differences were greater in DMDch than in controls for both parameters.ConclusionsThe myocardium in DMDch displays UTC features different from those in healthy controls. These results show that lack of dystrophin is commonly associated with changes in myocardial features well before the onset of changes of systolic function and overt cardiomyopathy
Conversion of nanoscale topographical information of cluster-assembled zirconia surfaces into mechanotransductive events promotes neuronal differentiation
Additional file 4: Table S1. Proteomic data for upregulated proteins. Proteins upregulated (compared to flat-Zr) or present only in cells grown on ns-Zr15. Adhesome proteins and proteins with roles in mechanobiological processes are marked in dark and light grey, respectively
Standardized nanomechanical atomic force microscopy procedure (SNAP) for measuring soft and biological samples
We present a procedure that allows a reliable determination of the elastic (Young's) modulus of soft samples, including living cells, by atomic force microscopy (AFM). The standardized nanomechanical AFM procedure (SNAP) ensures the precise adjustment of the AFM optical lever system, a prerequisite for all kinds of force spectroscopy methods, to obtain reliable values independent of the instrument, laboratory and operator. Measurements of soft hydrogel samples with a well-defined elastic modulus using different AFMs revealed that the uncertainties in the determination of the deflection sensitivity and subsequently cantilever's spring constant were the main sources of error. SNAP eliminates those errors by calculating the correct deflection sensitivity based on spring constants determined with a vibrometer. The procedure was validated within a large network of European laboratories by measuring the elastic properties of gels and living cells, showing that its application reduces the variability in elastic moduli of hydrogels down to 1%, and increased the consistency of living cells elasticity measurements by a factor of two. The high reproducibility of elasticity measurements provided by SNAP could improve significantly the applicability of cell mechanics as a quantitative marker to discriminate between cell types and conditions
Large expert-curated database for benchmarking document similarity detection in biomedical literature search
Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe
Imidazolium-Based Ionic Liquids Affect Morphology and Rigidity of Living Cells: an Atomic Force Microscopy Study
The study of the toxicity, biocompatibility, and environmental sustainability of room-temperature Ionic Liquids (ILs) is still in its infancy. Understanding the impact of ILs on living organisms, especially from the aquatic ecosystem, is urgent, since on one side large amounts of these substances are widely employed as solvents in industrial chemical processes, and on the other side evidences of toxic effects of ILs on microorganisms and single cells have been observed. To date, the toxicity of ILs have been investigated by means of macroscopic assays aimed at characterizing the effective concentrations (like the EC50) that cause the dead of a significant fraction of the population of microorganisms and cells. These studies allowed to identify the cell membrane as the first target of the IL interaction, whose effectiveness was correlated to the lipophilicity of the cation, i.e. to the length of the lateral alkyl chain. Our study aimed at characterizing the molecular mechanisms of the toxicity of ILs. To this purpose, we carried out a combined topographic and mechanical analysis by Atomic Force Microscopy of living breast metastatic cancer cells (MDA-MB-231) upon interaction with imidazolium-based ILs. We showed that ILs are able to induce modifications of the overall rigidity (effective Young modulus) and morphology of the cells. Our results demonstrate that ILs act on the physical properties of the cell membrane, and possibly induce cytoskeletal reorganization, already at concentrations below the EC50. These potentially toxic effects are stronger at higher IL concentrations, as well as with longer lateral chains in the cation.<br /
Surface confinement induces the formation of solid - like insulating ionic liquid nanostructures
We report on the modification of the electric properties of the imidazolium-based [BMIM][NTf2] ionic liquid upon surface confinement in the sub-monolayer regime. Solid-like insulating nanostructures of [BMIM][NTf2] spontaneously form on a variety of insulating substrates, at odd with the liquid and conductive nature of the same substances in the bulk phase. A systematic spatially-resolved investigation by atomic force microscopy of the morphological, mechanical and electrical properties of [BMIM][NTf2] nanostructures showed that this liquid substance rearranges into lamellar nanostructures with a high degree of vertical order and enhanced resistance to mechanical compressive stresses and very intense electric fields, denoting a solid-like character. The morphological and structural reorganization has a profound impact on the electric properties of supported [BMIM][NTf2] islands, which behave like insulator layers with a relative dielectric constant between 3 and 5, comparable to those of conventional ionic solids, and significantly smaller than those measured in the bulk ionic liquid. These results suggest that in the solid-like ordered domains confined either at surfaces or inside the pores of the nanoporous electrodes of photo-electrochemical devices, the ionic mobility and the overall electrical properties can be significantly perturbed with respect to the bulk liquid phase, which would likely influence the performance of the devices.<br /
The Effect of a DC Magnetic Field on the AC Magnetic Properties of Oleic Acid-Coated Fe<sub>3</sub>O<sub>4</sub> Nanoparticles
The AC magnetic properties of a sample of Fe3O4 nanoparticles coated with oleic acid have been investigated with the help of AC susceptibility measurements. In particular, several DC magnetic fields have been superimposed on the AC field, and their effect on the magnetic response of the sample has been analysed. The results show the presence of a double peak structure in the imaginary component of the complex AC susceptibility measured as a function of the temperature. A preliminary evaluation of the Mydosh parameter for both peaks gives the information that each one of them is associated with a different state of interaction between nanoparticles. The two peaks evolve both in amplitude and position when the intensity of the DC field is changed. The field dependence of the peak position shows two different trends, and it is possible to study them in the framework of the currently existing theoretical models. In particular, a model of non-interacting magnetic nanoparticles has been used to describe the behaviour of the peak at lower temperatures, whereas the behaviour of the peak at higher temperatures has been analysed in the framework of a spin-glass-like model. The proposed analysis technique can be useful for the characterisation of magnetic nanoparticles used in several types of applications, such as biomedical and magnetic fluids
Effect of Magnetite Nanoparticles Content on the Magnetic Properties of Polylactide and Polystyrene Composites
The effect of magnetite nanoparticles (NPs) content on the magnetic properties of polylactide (PLA) and polystyrene (PS) matrix has been investigated by means of DC magnetization measurements as a function of temperature (T) and magnetic field (H). Previous to the dispersion by melt-compounding into PLA and PS, the magnetite NPs have been reactively surface treated with 3% polymethylhydrogensiloxane (MHX) in order to make them hydrophobic and more stable to the action of oxygen and moisture. The magnetic analysis of the properties has been performed by measuring the Zero Field Cooling (ZFC) magnetization curve as a function of the temperature, at 0.1 Tesla applied field. In this framework, a superparamagnetic shape like has been noted for all the samples with the possibility to individuate the blocking temperature (TB) of the NPs. Moreover, the magnetization as a function of the field has been measured at room temperature (in particular above TB) investigating the coercive field and the magnetization values finding potentially interesting results. In particular, the very low values obtained for the coercive field at room temperature, together with the maximum found in the ZFC curve, have confirmed the superparamagnetic behavior of the PLA and PS - magnetite filled samples. Finally, a table with the fundamental magnetic features values of the samples has been reported for understanding if the obtained results for this kind of surface treated NPs and the afferent nanocomposites could be suitable for being used in applications requiring superparamagnetic properties (protection of environment, magnetic microcarriers, magnetic separation of stem cells, other biomedical purposes)
Cells nanomechanics by atomic force microscopy: focus on interactions at nanoscale
Nanomechanics of cytoskeleton is deeply involved in physiology and regulation of cell behavior. Atomic Force Microscopy has been extensively used for quantitative characterization with high-spatial resolution, in particular showing tremendous opportunities in biomechanics by quantifying mechanical parameters related to cytoskeleton organization. In this short review, we highlight recent developments in cell nanomechanics by AFM focusing on methodology and direct application to investigate cytoskeleton restructuration when cells are interacting with nanostructures (surfaces and nanoparticles). In particular, cells can sense the stiffness of environment or internalized particles and AFM can detect the rearrangement of cytoskeleton as one of the responses of mechanotransduction stimuli. Current bottlenecks hindering further progress in technology, such as theoretical models of interpretation will be discussed, in particular we propose a solution for complex system by coupling AFM with finite element simulations to retrieve more quantitative information when heterogeneity and convolution play important roles. Finally, we present recent cutting-edge research directions to explore new techniques and enhance the capabilities of AFM nanomechanics for living cells
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