110 research outputs found
Direct measurement of spatial modes of a micro-cantilever from thermal noise
International audienceMeasurements of the deflection induced by thermal noise have been performed on a rectangular atomic force microscope cantilever in air. The detection method, based on polarization interferometry, can achieve a resolution of 1E-14 m/rtHz in the frequency range 1 kHz – 800 kHz. The focused beam from the interferometer probes the cantilever at different positions along its length and the spatial modes' shapes are determined up to the fourth resonance, without external excitation. Results are in good agreement with theoretically expected behavior. From this analysis accurate determination of the elastic constant of the cantilever is also achieved
Nonlinear behaviour of self-excited microcantilevers in viscous fluids
Microcantilevers are increasingly being used to create sensitive sensors for rheology and mass sensing at the micro- and nano-scale. When operating in viscous liquids, the low quality factor of such resonant structures, translating to poor signal-to-noise ratio, is often manipulated by exploiting feedback strategies. However, the presence of feedback introduces poorly-understood dynamical behaviours that may severely degrade the sensor performance and reliability. In this paper, the dynamical behaviour of self-excited microcantilevers vibrating in viscous fluids is characterized experimentally and two complementary modelling approaches are proposed to explain and predict the behaviour of the closed-loop system. In particular, the delay introduced in the feedback loop is shown to cause surprising non-linear phenomena consisting of shifts and sudden-jumps of the oscillation frequency. The proposed dynamical models also suggest strategies for controlling such undesired phenomena
Measuring Viscosity Using the Hysteresis of the Non-Linear Response of a Self-Excited Cantilever
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Self-driven soft imaging in liquid by means of photothermal excitation
The use of a laser to induce oscillations of an atomic force microscopy cantilever provides a way to excite the dynamics of the system in a very controlled manner. This excitation scheme has been used to obtain reliable self-sustained oscillations, in air and in liquid environments, and to implement an additional control loop leading to a soft, low-interaction, working mode. The dynamics of the oscillating cantilever has been characterized, both theoretically and experimentally, and the effectiveness of the approach has been verified on a test sample. © 2011 American Institute of Physics.Peer Reviewe
Catechol-Containing Hydroxylated Biomimetic 4-Thiaflavanes as Inhibitors of Amyloid Aggregation
The study of compounds able to interfere in various ways with amyloid aggregation is of paramount importance in amyloid research. Molecules characterized by a 4-thiaflavane skeleton have received great attention in chemical, medicinal, and pharmaceutical research. Such molecules, especially polyhydroxylated 4-thiaflavanes, can be considered as structural mimickers of several natural polyphenols that have been previously demonstrated to bind and impair amyloid fibril formation. In this work, we tested five different 4-thiaflavanes on the hen egg-white lysozyme (HEWL) amyloid model for their potential anti-amyloid properties. By combining a thioflavin T assay, atomic force microscopy, and a cell toxicity assay, we demonstrated that such compounds can impair the formation of high-order amyloid aggregates and mature fibrils. Despite this, the tested 4-thiaflavanes, although non-toxic per se, are not able to prevent amyloid toxicity on human neuroblastoma cells. Rather, they proved to block early aggregates in a stable, toxic conformation. Accordingly, 4-thiaflavanes can be proposed for further studies aimed at identifying blocking agents for the study of toxicity mechanisms of amyloid aggregation
Mechanisms for the inhibition of amyloid aggregation by small ligands
The formation of amyloid aggregates is the hallmark of systemic and neurodegenerative disorders, also known as amyloidoses. Many proteins have been found to aggregate into amyloid-like fibrils and this process is recognized as a general tendency of polypeptides. Lysozyme, an antibacterial protein, is a well-studied model since it is associated in human with systemic amyloidosis and that is widely available from chicken eggs (HEWL, hen egg white lysozyme). In the present study we investigated the mechanism of interaction of aggregating HEWL with rosmarinic acid and resveratrol, that we verified to be effective and ineffective, respectively, in inhibiting aggregate formation. We used a multidisciplinary strategy to characterize such effects, combining biochemical and biophysical methods with molecular dynamics (MD) simulations on the HEWL peptide 49–64 to gain insights into the mechanisms and energy variations associated to amyloid formation and inhibition. MD revealed that neither resveratrol nor rosmarinic acid were able to compete with the initial formation of the β-sheet structure. We then tested the association of two β-sheets, representing the model of an amyloid core structure. MD showed that rosmarinic acid displayed an interaction energy and a contact map comparable to that of sheet pairings. On the contrary, resveratrol association energy was found to be much lower and its contact map largely different than that of sheet pairings. The overall characterization elucidated a possible mechanism explaining why, in this model, resveratrol is inactive in blocking fibril formation, whereas rosmarinic acid is instead a powerful inhibitor
Opto-mechanical probe for combining atomic force microscopy and optical near-field surface analysis
We have developed a new easy-to-use probe that can be used to combine atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM). We show that, using this device, the evanescent field, obtained by total internal reflection conditions in a prism, can be visualized by approaching the surface with the scanning tip. Furthermore, we were able to obtain simultaneous AFM and SNOM images of a standard test grating in air and in liquid. The lateral resolution in AFM and SNOM mode was estimated to be 45 and 160 nm, respectively. This new probe overcomes a number of limitations that commercial probes have, while yielding the same resolution. (C) 2014 Optical Society of Americ
The induction of α-helical structure in partially unfolded HypF-N does not affect its aggregation propensity
The conversion of proteins into structured fibrillar aggregates is a central problem in protein chemistry, biotechnology, biology and medicine. It is generally accepted that aggregation takes place from partially structured states of proteins. However, the role of the residual structure present in such conformational states is not yet understood. In particular, it is not yet clear as to whether the α-helical structure represents a productive or counteracting structural element for protein aggregation. We have addressed this issue by studying the aggregation of pH-unfolded HypF-N. It has previously been shown that the two native α-helices of HypF-N retain a partial α-helical structure in the pH-unfolded state and that these regions are also involved in the formation of the cross-β structure of the aggregates. We have introduced mutations in such stretches of the sequence, with the aim of increasing the α-helical structure in the key regions of the pH-unfolded state, while minimizing the changes of other factors known to influence protein aggregation, such as hydrophobicity, β-Sheet propensity, etc. The resulting HypF-N mutants have higher contents of α-helical structure at the site(s) of mutation in their pH-unfolded states, but such an increase does not correlate with a change of aggregation rate. The results suggest that stabilisation of α-helical structure in amyloidogenic regions of the sequence of highly dynamic states does not have remarkable effects on the rate of protein aggregation from such conformational states. Comparison with other protein systems indicate that the effect of increasing α-helical propensity can vary if the stabilised helices are in non-amyloidogenic stretches of initially unstructured peptides (accelerating effect), in amyloidogenic stretches of initially unstructured peptides (no effect) or in amyloidogenic stretches of initially stable helices (decelerating effect
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