Conductive Atomic Force Microscopy-Ultralow-Current Measurement Systems for Nanoscale Imaging of a Surface's Electrical Properties.

One of the most advanced and versatile nanoscale diagnostic tools is atomic force microscopy. By enabling advanced imaging techniques, it allows us to determine various assets of a surface, including morphological, electrical, mechanical, magnetic, and thermal properties. Measuring local current flow is one of the very important methods of evaluation for, for instance, photovoltaic materials or semiconductor structures and other nanodevices. Due to contact areas, the current densities can easily reach above 1 kA/m2; therefore, special detection/measurement setups are required. They meet the required measurement range, sensitivity, noise level, and bandwidth at the measurement scale. Also, they prevent the sample from becoming damaged and prevent unwanted tip-sample issues. In this paper, we present three different nanoscale current measurement solutions, supported with test results, proving their performance.

Influence of Glycerol on the Surface Morphology and Crystallinity of Polyvinyl Alcohol Films.

The structure and physicochemical properties of polyvinyl alcohol (PVA) and PVA/glycerol films have been investigated by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), thermogravimetry/differential thermal analysis (TG/DTA), and advanced scanning probe microscopy (SPM). In the pure PVA films, SPM allowed us to observe ribbon-shaped domains with a different frictional and elastic contrast, which apparently originated from a correlated growth or assembly of PVA crystalline nuclei located within individual PVA clusters. The incorporation of 22% w/w glycerol led to modification in shape of those domains from ribbon-like in pure PVA to rounded in PVA/glycerol 22% w/w films; changes in the relative intensities of the XRD peaks and a decrease in the amorphous halo in the XRD pattern were also detected, while the DTA peak corresponding to the melting point remained at almost the same temperature. For higher glycerol content, FT-IR revealed additional glycerol-characteristic peaks presumably related to the formation of glycerol aggregates, and XRD, FT-IR, and DTA all indicated a reduction in crystallinity. For more than 36% w/w glycerol, the plasticization of the films complicated the acquisition of SPM images without tip-induced surface modification. Our study contributes to the understanding of crystallinity in PVA and how it is altered by a plasticizer such as glycerol.

Evaluating Optical Properties of Mixed-Phase 2D MoSe2/Poly(vinyl alcohol) Nanocomposite Film.

Highly solar light-absorbing poly(vinyl alcohol) (PVA) nanocomposite films have garnered wide attention in fields such as flexible optoelectronics, solar energy harvesting, and photothermal therapy. However, fabricating PVA nanocomposite films with a broad spectrum of solar absorption using cost-effective and non-toxic nanofillers remains challenging. Herein, nanocomposite films of PVA incorporating various concentrations of mixed-phase 2D MoSe2 nanosheets (i.e., a combination of the 2H and 1T phase) were prepared using a solution casting technique. Scanning electron microscopy (SEM) shows homogenous dispersion of MoSe2 nanosheets in the PVA matrix even at higher concentrations, while atomic force microscopy (AFM) reveals increasing surface roughness with increasing MoSe2 content, reaching a plateau after 20 wt%. With the increase in the concentration of MoSe2, the nanocomposite films exhibit interesting light absorption characteristics reaching their highest absorption (average 94.9%) at 40 wt% MoSe2. The incorporated mixed-phase MoSe2 nanosheets induce a significant change in the energy levels of the PVA matrix, which is reflected in the reduced optical band gap energy (2.63 eV) at 40 wt% MoSe2 against pure PVA (5.28 eV). The excellent light absorption of PVA nanocomposite films across the entire range from 250 nm to 2500 nm is attributed to the thin 2D structure of MoSe2 and the presence of its mixed phase.

Effects of nanoparticle deformability on multiscale biotransport.

Deformability is one of the critical attributes of nanoparticle (NP) drug carriers, along with size, shape, and surface properties. It affects various aspects of NP biotransport, ranging from circulation and biodistribution to interactions with biological barriers and target cells. Recent studies report additional roles of NP deformability in biotransport processes, including protein corona formation, intracellular trafficking, and organelle distribution. This review focuses on the literature published in the past five years to update our understanding of NP deformability and its effect on NP biotransport. We introduce different methods of modulating and evaluating NP deformability and showcase recent studies that compare a series of NPs in their performance in biotransport events at all levels, highlighting the consensus and disagreement of the findings. It concludes with a perspective on the intricacy of systematic investigation of NP deformability and future opportunities to advance its control toward optimal drug delivery.

Investigation on Junction Contacts of Semiconducting Carbon Nanotube Networks Using Conductive Atomic Force Microscopy.

Semiconductor single-walled carbon nanotube (s-SWNT) networks have gained prominence in electronic devices due to their cost-effectiveness, relatively production-naturality, and satisfactory performance. Configuration, density, and resistance of SWNT-SWNT junctions are considered crucial factors influencing the overall conductivity of s-SWNT networks. In this study, we present a method for inferring the lower bounds of the SWNT-SWNT junction resistance in s-SWNT networks based on conductive atomic force microscopy TUNA images. This method further enables the proposal of a classification for SWNT-SWNT junctions based on the current behavior relative to their surroundings. The three types of SWNT-SWNT junctions are denoted as (i) true contact (T), (ii) poor contact (P), and (iii) false contact (F). Of them, the true and poor contacts, respectively, represent good and poor electrical contact for the subject SWNT-SWNT junctions whose electrical conductivity hardly improves under external tip pressure, while that of the false contact can be further improved by external pressure. Statistical analysis demonstrates that while T-type junctions make a significant contribution to network conductivity, their proportion accounts for only approximately 40%. The P-type and F-type junctions, which constitute over 60% of the total, may be a contributing factor that constrains the overall conductivity of the s-SWNT networks. The height ratio of the junction to the sum of two SWNTs was also observed to exhibit variations among the three types. Finally, we propose a three-dimensional model to elucidate the formation mechanism underlying each type of junction. The present study provides insights into the performance of spontaneous contacts between s-SWNTs in the networks, and the systematic image acquisition and junction classification processes may provide support for future advancements in these networks.

Effect of the anti-Alzheimer drug GSK-3ß antagonist on numerical modeling of the energy dissipation through the resin-dentin interface.

OBJECTIVES: The aim of this study was to determine the viscoelastic performance and energy dissipation of conditioned dentin infiltrated with polymeric nanoparticles (NPs) doped with tideglusib (TDg) (TDg-NPs). METHODS: Dentin conditioned surfaces were infiltrated with NPs and TDg-NPs. Bonded interfaces were created, stored for 24 h and submitted to mechanical and thermal challenging. Resin-dentin interfaces were evaluated through nano-DMA/complex-loss-storage moduli-tan delta assessment and atomic force microscopy (AFM) analysis. RESULTS: Dentin infiltrated with NPs and load cycled attained the highest complex modulus at hybrid layer and bottom of hybrid layer. Intertubular dentin treated with undoped NPs showed higher complex modulus than peritubular dentin, after load cycling, provoking energy concentration and breakdown at the interface. After infiltrating with TDg-NPs, complex modulus was similar between peri-intertubular dentin and energy dissipated homogeneously. Tan delta at intertubular dentin was higher than at peritubular dentin, after using TDg-NPs and load cycling. This generated the widest bandwidth of the collagen fibrils and bridge-like mineral structures that, as sight of energy dissipation, fastened active dentin remodeling. TDg-NPs inducted scarce mineralization after thermo-cycling, but these bridging processes limited breakdown zones at the interface. SIGNIFICANCE: TDg-based NPs are then proposed for effective dentin remineralization and tubular seal, from a viscoelastic approach.

Methylglyoxal alters collagen fibril nanostiffness and surface potential.

Collagen fibrils are fundamental to the mechanical strength and function of biological tissues. However, they are susceptible to changes from non-enzymatic glycation, resulting in the formation of advanced glycation end-products (AGEs) that are not reversible. AGEs accumulate with aging and disease and can adversely impact tissue mechanics and cell-ECM interactions. AGE-crosslinks have been related, on the one hand, to dysregulation of collagen fibril stiffness and damage and, on the other hand, to altered collagen net surface charge as well as impaired cell recognition sites. While prior studies using Kelvin probe force microscopy (KPFM) have shown the effect glycation has on collagen fibril surface potential (i.e., net charge), the combined effect on individual and isolated collagen fibril mechanics, hydration, and surface potential has not been documented. Here, we explore how methylglyoxal (MGO) treatment affects the mechanics and surface potential of individual and isolated collagen fibrils by utilizing atomic force microscopy (AFM) nanoindentation and KPFM. Our results reveal that MGO treatment significantly increases nanostiffness, alters surface potential, and modifies hydration characteristics at the collagen fibril level. These findings underscore the critical impact of AGEs on collagen fibril physicochemical properties, offering insights into pathophysiological mechanical and biochemical alterations with implications for cell mechanotransduction during aging and in diabetes. STATEMENT OF SIGNIFICANCE: Collagen fibrils are susceptible to glycation, the irreversible reaction of amino acids with sugars. Glycation affects the mechanical properties and surface chemistry of collagen fibrils with adverse alterations in biological tissue mechanics and cell-ECM interactions. Current research on glycation, at the level of individual collagen fibrils, is sparse and has focused either on collagen fibril mechanics, with contradicting evidence, or surface potential. Here, we utilized a multimodal approach combining Kelvin probe force (KPFM) and atomic force microscopy (AFM) to examine how methylglyoxal glycation induces structural, mechanical, and surface potential changes on the same individual and isolated collagen fibrils. This approach helps inform structure-function relationships at the level of individual collagen fibrils.

Pulmonary surfactant and prostaglandin E2 in airway smooth muscle relaxation of human and male guinea pigs.

Pulmonary surfactant serves as a barrier to respiratory epithelium but can also regulate airway smooth muscle (ASM) tone. Surfactant (SF) relaxes contracted ASM, similar to ß2-agonists, anticholinergics, nitric oxide, and prostanoids. The exact mechanism of surfactant relaxation and whether surfactant relaxes hyperresponsive ASM remains unknown. Based on previous research, relaxation requires an intact epithelium and prostanoid synthesis. We sought to examine the mechanisms by which surfactant causes ASM relaxation. Organ bath measurements of isometric tension of ASM of guinea pigs in response to exogenous surfactant revealed that surfactant reduces tension of healthy and hyperresponsive tracheal tissue. The relaxant effect of surfactant was reduced if prostanoid synthesis was inhibited and/or if prostaglandin E2-related EP2 receptors were antagonized. Atomic force microscopy revealed that human ASM cells stiffen during contraction and soften during relaxation. Surfactant softened ASM cells, similarly to the known bronchodilator prostaglandin E2 (PGE2) and the cell softening was abolished when EP4 receptors for PGE2 were antagonized. Elevated levels of PGE2 were found in cultures of normal human bronchial epithelial cells exposed to pulmonary surfactant. We conclude that prostaglandin E2 and its EP2 and EP4 receptors are likely involved in the relaxant effect of pulmonary surfactant in airways.

FabR, a regulator of membrane lipid homeostasis, is involved in Klebsiella pneumoniae biofilm robustness.

Biofilm is a dynamic structure from which individual bacteria and micro-aggregates are released to subsequently colonize new niches by either detachment or dispersal. Screening of a transposon mutant library identified genes associated with the alteration of Klebsiella pneumoniae biofilm including fabR, which encodes a transcriptional regulator involved in membrane lipid homeostasis. An isogenic ∆fabR mutant formed more biofilm than the wild-type (WT) strain and its trans-complemented strain. The thick and round aggregates observed with ∆fabR were resistant to extensive washes, unlike those of the WT strain. Confocal microscopy and BioFlux microfluidic observations showed that fabR deletion was associated with biofilm robustness and impaired erosion over time. The genes fabB and yqfA associated with fatty acid metabolism were significantly overexpressed in the ∆fabR strain, in both planktonic and biofilm conditions. Two monounsaturated fatty acids, palmitoleic acid (C16:1) and oleic acid (C18:1), were found in higher proportion in biofilm cells than in planktonic forms, whereas heptadecenoic acid (C17:1) and octadecanoic acid, 11-methoxy (C18:0-OCH3) were found in higher proportion in the planktonic lifestyle. The fabR mutation induced variations in the fatty acid composition, with no clear differences in the amounts of saturated fatty acids (SFA) and unsaturated fatty acids for the planktonic lifestyle but lower SFA in the biofilm form. Atomic force microscopy showed that deletion of fabR is associated with decreased K. pneumoniae cell rigidity in the biofilm lifestyle, as well as a softer, more elastic biofilm with increased cell cohesion compared to the wild-type strain.IMPORTANCEKlebsiella pneumoniae is an opportunistic pathogen responsible for a wide range of nosocomial infections. The success of this pathogen is due to its high resistance to antibiotics and its ability to form biofilms. The molecular mechanisms involved in biofilm formation have been largely described but the dispersal process that releases individual and aggregate cells from mature biofilm is less well documented while it is associated with the colonization of new environments and thus new threats. Using a multidisciplinary approach, we show that modifications of bacterial membrane fatty acid composition lead to variations in the biofilm robustness, and subsequent bacterial detachment and biofilm erosion over time. These results enhance our understanding of the genetic requirements for biofilm formation in K. pneumoniae that affect the time course of biofilm development and the embrittlement step preceding its dispersal that will make it possible to control K. pneumoniae infections.

Quantitative study of early-stage transient bacterial adhesion to bioactive glass and glass ceramics: atomic force microscopic observations.

Antimicrobial potential of bioactive glass (BAG) makes it promising for implant applications, specifically overcoming the toxicity concerns associated with traditional antibacterial nanoparticles. The 58S composition of BAG (with high Ca and absence of Na) has been known to exhibit excellent bioactivity and antibacterial behaviour, but the mechanisms behind have not been investigated in detail. In this pioneering study, we are using Atomic Force Microscopy (AFM) to gain insights into 58S BAG's adhesive interactions with planktonic cells of both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria; along with the impact of crystallinity on antibacterial properties. We have recorded greater bacterial inhibition by amorphous BAG compared to semi-crystalline glass-ceramics and stronger effect against gram-negative bacteria via conventional long-term antibacterial tests. AFM force distance curves has illustrated substantial bonding between bacteria and BAG within the initial one second (observed at a gap of 250 ms) of contact, with multiple binding events. Further, stronger adhesion of BAG with E.coli (~ 6 nN) compared to S. aureus (~ 3 nN) has been found which can be attributed to more adhesive nano-domains (size effect) distributed uniformly on E.coli surface. This study has revealed direct evidence of impact of contact time and 58S BAG's crystalline phase on bacterial adhesion and antimicrobial behaviour. Current study has successfully demonstrated the mode and mechanisms of initial bacterial adhesion with 58S BAG. The outcome can pave the way towards improving the designing of implant surfaces for a range of biomedical applications.

Broad Adaptability of Coronavirus Adhesion Revealed from the Complementary Surface Affinity of Membrane and Spikes.

Coronavirus stands for a large family of viruses characterized by protruding spikes surrounding a lipidic membrane adorned with proteins. The present study explores the adhesion of transmissible gastroenteritis coronavirus (TGEV) particles on a variety of reference solid surfaces that emulate typical virus-surface interactions. Atomic force microscopy informs about trapping effectivity and the shape of the virus envelope on each surface, revealing that the deformation of TGEV particles spans from 20% to 50% in diameter. Given this large deformation range, experimental Langmuir isotherms convey an unexpectedly moderate variation in the adsorption-free energy, indicating a viral adhesion adaptability which goes beyond the membrane. The combination of an extended Helfrich theory and coarse-grained simulations reveals that, in fact, the envelope and the spikes present complementary adsorption affinities. While strong membrane-surface interaction lead to highly deformed TGEV particles, surfaces with strong spike attraction yield smaller deformations with similar or even larger adsorption-free energies.

Evaluation of hair surface structure and morphology of patients with lichen planopilaris (LPP) by atomic force microscopy (AFM).

BACKGROUND: Lichen planopilaris (LPP) is a chronic lymphocytic skin disease manifested by progressive scarring alopecia. The diagnosis of LPP is made based on histopathological examination, although it is not always definite. The current study evaluates the effectiveness of non-invasive atomic force microscopy (AFM) hair examination in detecting morphological differences between healthy and diseased hair. MATERIALS AND METHODS: Here, three to five hairs from lesional skin of 10 LPP patients were collected and examined at nine locations using AFM. At least four images were taken at each of the nine sites. Metric measurements were taken and metric (length, width, and scale step height) and morphological features (striated and smooth surface of scales, the presence of endocuticle and cortex, shape of scales edges, scratches, pitting, cracks, globules, and wavy edge) were compared with hair from healthy controls. In addition, areas on diseased hair where the process of pathological, unnatural delamination of the hair fiber occurs are described. RESULTS: There was a statistically significant difference in the number of scratches in the initial sections of the LPP hair, in the intensity of wavy edges along the entire length of the tested hair, and in the number of scales with pitting in the middle section of the hair. In addition, a statistically significant higher number of scales with striated surface was found in LPP group starting at 3.5 cm from the root continuing towards the free end of the hair. Other morphological changes such as presence of cortex, globules, oval indentations, and rod-like macrofibrillar elements were also assessed, however, detailed results are not presented, as the differences shown in the number of these morphological changes were not significantly different. CONCLUSION: This publication outlines the differences between virgin, healthy Caucasian hair, and the hair of LPP patients. The results of this study can be used for further research and work related to LPP. This is the first attempt to characterize the hair of LPP patients using AFM.

Atomic force microscopy as a nanomechanical tool for cancer liquid biopsy.

Liquid biopsies have been receiving tremendous attention for their potential to reshape cancer management. Though current studies of cancer liquid biopsy primarily fucus on applying biochemical assays to characterize the genetic/molecular profiles of circulating tumor cells (CTCs) and their secondary products shed from tumor sites in bodily fluids, delineating the nanomechanical properties of tumor-associated materials in liquid biopsy specimens yields complementary insights into the biology of tumor dissemination and evolution. Particularly, atomic force microscopy (AFM) has become a standard and versatile toolbox for characterizing the mechanical properties of living biological systems at the micro/nanoscale, and AFM has been increasingly utilized to probe the nanomechanical properties of various tumor-derived analytes in liquid biopsies, including CTCs, tumor-associated cells, circulating tumor DNA (ctDNA) molecules, and extracellular vesicles (EVs), offering additional possibilities for understanding cancer pathogenesis from the perspective of mechanobiology. Herein, the applications of AFM in cancer liquid biopsy are summarized, and the challenges and future directions of AFM as a nanomechanical analysis tool in cancer liquid biopsy towards clinical utility are discussed and envisioned.

Investigation of Mode-Induced Spin Wave Transmission Blockage by In Situ Nanoscale Grooves.

In the pursuit of advancing spin-wave optics, the propagation of magnetostatic surface spin-waves is investigated in a uniform permalloy waveguide with in-situ nanopatterned grooves created through Atomic Force Microscopy nanolithography and Focused Ion Beam etching. The study unveils that the introduction of narrow constrictions and grooves leads to a non-monotonic reduction of the transmitted spin-wave signal intensity as the spin-wave pathway is shrinked. The remarkable feature that a stronger signal extinction is obtained for a narrow groove compared to a spin-waveguide interrupted by a full gap, where only inefficient transport through dipolar coupling is allowed, is highlighted. Combining experimental and numerical analyses, the intricate interplay between spin-wave diffraction and reflection at the waveguide edges is unraveled, being at the origin of a transverse-mode variation responsible for the signal extinction when detected using coplanar antennas. The findings offer insights into the controllable manipulation of detected spin-wave intensity, thereby opening promising avenues for the improvement of spin-wave switches and interferometers, and for the nanopatterning of graded index magnonics.

The Effect of Relative Humidity in Conductive Atomic Force Microscopy.

Conductive atomic force microscopy (CAFM) analyzes electronic phenomena in materials and devices with nanoscale lateral resolution, and it is widely used by companies, research institutions, and universities. Most data published in the field of CAFM is collected in air at a relative humidity (RH) of 30-60%. However, the effect of RH in CAFM remains unclear because previous studies often made contradictory claims, plus the number of samples and locations tested is scarce. Moreover, previous studies on this topic did not apply current limitations, which can degrade the CAFM tips and generate false data. This article systematically analyzes the effect of RH in CAFM by applying ramped voltage stresses at over 17,000 locations on ten different samples (insulating, semiconducting, and conducting) under seven different RH. An ultra-reliable setup with a 110-pA current limitation during electrical stresses is employed, and excellent CAFM tip integrity after thousands of tests is demonstrated. It is found that higher RH results in increased currents due to the presence of a conductive water meniscus at the tip/sample junction, which increases the effective area for electron flow. This trend is observed in insulators and ultra-thin semiconductors; however, in thicker semiconductors the electron mean free path is high enough to mask this effect. Metallic samples show no dependence on RH. This study clarifies the effect of relative humidity in CAFM, enhances understanding of the technique, and teaches researchers how to improve the reliability of their studies in this field.

(1-Deoxy)ceramides in bilayers containing sphingomyelin and cholesterol.

The discovery of a novel sphingolipid subclass, the (1-deoxy)sphingolipids, which lack the 1-hydroxy group, attracted considerable attention in the last decade, mainly due to their involvement in disease. They differed in their physico-chemical properties from the canonical (or 1-hydroxy) sphingolipids and they were more toxic when accumulated in cells, inducing neurodegeneration and other dysfunctions. (1-Deoxy)ceramides, (1-deoxy)dihydroceramides, and (1- deoxymethyl)dihydroceramides, the latter two containing a saturated sphingoid chain, have been studied in this work using differential scanning calorimetry, confocal fluorescence and atomic force microscopy, to evaluate their behavior in bilayers composed of mixtures of three or four lipids. When compared to canonical ceramides (Cer), a C16:0 (1-deoxy)Cer shows a lower miscibility in mixtures of the kind C16:0 sphingomyelin/cholesterol/XCer, where XCer is any (1-deoxy)ceramide, giving rise to the coexistence of a liquid-ordered phase and a gel phase. The latter resembles, in terms of thermotropic behavior and nanomechanical resistance, the gel phase of the C16:0 sphingomyelin/cholesterol/C16:0 Cer mixture [Busto et al., Biophys. J. 2014, 106, 621-630]. Differences are seen between the various C16:0 XCer under study in terms of nanomechanical resistance, bilayer thickness and bilayer topography. When examined in a more fluid environment (bilayers based on C24:1 SM), segregated gel phases are still present. Probably related to such lateral separation, XCer preserve the capacity for membrane permeation, but their effects are significantly lower than those of canonical ceramides. Moreover, C24:1 XCer show significantly lower membrane permeation capacity than their C16:0 counterparts. The above data may be relevant in the pathogenesis of certain sphingolipid-related diseases, including certain neuropathies, diabetes, and glycogen storage diseases.

Surface Degradation of Mg2X-Based Composites at Room Temperature: Assessing Grain Boundary and Bulk Diffusion Using Atomic Force Microscopy and Scanning Electron Microscopy.

Practical application of thermoelectric generators necessitates materials that combine high heat-to-electricity conversion efficiency with long-term functional stability under operation conditions. While Mg2(Si,Sn)-based materials exhibit promising thermoelectric properties and module prototypes have been demonstrated, their stability remains a challenge, demanding thorough investigation. Utilizing atomic force microscopy (AFM) and scanning electron microscopy (SEM), we investigate the surface degradation of a composite material comprising Si-rich and Sn-rich Mg2(Si,Sn) solid solutions. The investigation reveals a pronounced dependence of stability on Sn content, with the Sn-rich phase Mg2Si0.13Sn0.87 displaying the formation of a nonprotective oxide layer. Subsequent AFM measurements provide evidence of dominating grain boundary diffusion of loosely bound Mg, compared to bulk diffusion, observed within a few days, ultimately resulting in a complete surface oxidation of the Sn-rich phase within several weeks. On the other hand, Mg2Si and Si-rich Mg2Si0.80±0.05Sn0.20±0.05 remain stable against Mg diffusion to the surface even after prolonged exposure. Comparison with previous investigations confirms that the degradation rate is found to be highly dependent on the Sn content, with markedly higher rates observed for x = 0.87 compared to x = 0.70 in Mg2Si1-xSnx. These findings contribute to a better understanding of the stability challenges associated with Mg2(Si,Sn)-based materials, essential for the development of robust thermoelectric materials for practical applications.

Architecture of native kinetochores revealed by structural studies utilizing a thermophilic yeast.

Eukaryotic chromosome segregation requires kinetochores, multi-megadalton protein machines that assemble on the centromeres of chromosomes and mediate attachments to dynamic spindle microtubules. Kinetochores are built from numerous complexes, and there has been progress in structural studies on recombinant subassemblies. However, there is limited structural information on native kinetochore architecture. To address this, we purified functional, native kinetochores from the thermophilic yeast Kluyveromyces marxianus and examined them by electron microscopy (EM), cryoelectron tomography (cryo-ET), and atomic force microscopy (AFM). The kinetochores are extremely large, flexible assemblies that exhibit features consistent with prior models. We assigned kinetochore polarity by visualizing their interactions with microtubules and locating the microtubule binder, Ndc80c. This work shows that isolated kinetochores are more dynamic and complex than what might be anticipated based on the known structures of recombinant subassemblies and provides the foundation to study the global architecture and functions of kinetochores at a structural level.

Exploring the Challenges of Characterising Surface Topography of Polymer-Nanoparticle Composites.

Nanomechanical testing plays a crucial role in evaluating surfaces containing nanoparticles. Testing verifies surface performance concerning their intended function and detects any potential shortcomings in operational standards. Recognising that nanostructured surfaces are not always straightforward or uniform is essential. The chemical composition and morphology of these surfaces determine the end-point functionality. This can entail a layered surface using materials in contrast to each other that may require further modification after nanomechanical testing to pass performance and quality standards. Nanomechanical analysis of a structured surface consisting of a poly-methyl oxazoline film base functionalised with colloidal gold nanoparticles was demonstrated using an atomic force microscope (AFM). AFM nanomechanical testing investigated the overall substrate architecture's topographical, friction, adhesion, and wear parameters. Limitations towards its potential operation as a biomaterial were also addressed. This was demonstrated by using the AFM cantilever to apply various forces and break the bonds between the polymer film and gold nanoparticles. The AFM instrument offers an insight to the behaviour of low-modulus surface against a higher-modulus nanoparticle. This paper details the bonding and reaction limitations between these materials on the application of an externally applied force. The application of this interaction is highly scrutinised to highlight the potential limitations of a functionalised surface. These findings highlight the importance of conducting comprehensive nanomechanical testing to address concerns related to fabricating intricate biomaterial surfaces featuring nanostructures.

Modulation of arterial intima stiffness by disturbed blood flow.

The intima, comprising the endothelium and the subendothelial matrix, plays a crucial role in atherosclerosis pathogenesis. The mechanical stress arising from disturbed blood flow (d-flow) and the stiffening of the arterial wall contributes to endothelial dysfunction. However, the specific impacts of these physical forces on the mechanical environment of the intima remain undetermined. Here, we investigated whether inhibiting collagen crosslinking could ameliorate the detrimental effects of persistent d-flow on the mechanical properties of the intima. Partial ligation of the left carotid artery (LCA) was performed in C57BL/6J mice, inducing d-flow. The right carotid artery (RCA) served as an internal control. Carotids were collected 2 days and 2 weeks after surgery to study acute and chronic effects of d-flow on the mechanical phenotype of the intima. The chronic effects of d-flow were decoupled from the ensuing arterial wall stiffening by administration of ß-aminopropionitrile (BAPN), an inhibitor of collagen crosslinking by lysyl oxidase (LOX) enzymes. Atomic force microscopy (AFM) was used to determine stiffness of the endothelium and the denuded subendothelial matrix in en face carotid preparations. The stiffness of human aortic endothelial cells (HAEC) cultured on soft and stiff hydrogels was also determined. Acute exposure to d-flow caused a slight decrease in endothelial stiffness in male mice but had no effect on the stiffness of the subendothelial matrix in either sex. Regardless of sex, the intact endothelium was softer than the subendothelial matrix. In contrast, exposure to chronic d-flow led to a substantial increase in the endothelial and subendothelial stiffness in both sexes. The effects of chronic d-flow were largely prevented by concurrent BAPN administration. In addition, HAEC displayed reduced stiffness when cultured on soft vs. stiff hydrogels. We conclude that chronic d-flow results in marked stiffening of the arterial intima, which can be effectively prevented by inhibition of collagen crosslinking.