Distance dependence of the phase signal in eddy current microscopy

Atomic force microscopy using a magnetic tip is a promising tool for investigating conductivity on the nano-scale. By the oscillating magnetic tip eddy currents are induced in the conducting parts of the sample which can be detected in the phase signal of the cantilever. However, the origin of the phase signal is still controversial because theoretical calculations using a monopole appoximation for taking the electromagnetic forces acting on the tip into account yield an effect which is too small by more than two orders of magnitude. In order to determine the origin of the signal we used especially prepared gold nano patterns embedded in a non-conducting polycarbonate matrix and measured the distance dependence of the phase signal. Our data clearly shows that the interacting forces are long ranged and therefore, are likely due to the electromagnetic interaction between the magnetic tip and the conducting parts of the surface. Due to the long range character of the interaction a change in conductivity of $\Delta\sigma=4,5\cdot10^{7} (\Omega$m$)^{-1}$ can be detected far away from the surface without any interference from the topography

Anomalies in thickness measurements of graphene and few layer graphite crystals by tapping mode atomic force microscopy

Atomic Force Microscopy (AFM) in the tapping (intermittent contact) mode is a commonly used tool to measure the thickness of graphene and few layer graphene (FLG) flakes on silicon oxide surfaces. It is a convenient tool to quickly determine the thickness of individual FLG films. However, reports from literature show a large variation of the measured thickness of graphene layers. This paper is focused on the imaging mechanism of tapping mode AFM (TAFM) when measuring graphene and FLG thickness and we show that at certain measurement parameters significant deviations can be introduced in the measured thickness of FLG flakes. An increase of as much as 1 nm can be observed in the measured height of FLG crystallites, when using an improperly chosen range of free amplitude values of the tapping cantilever. We present comparative Raman spectroscopy and TAFM measurements on selected single and multilayer graphene films, based on which we suggest ways to correctly measure graphene and FLG thickness using TAFM

Conductive nanodots on the surface of irradiated CaF2

CaF2(111) single crystal surfaces have been irradiated with swift heavy ions under oblique angles resulting in chains of nanosized hillocks. In order to characterize these nanodots with respect to their conductivity we have applied non-contact atomic force microscopy using a magnetic tip. Measurements in UHV as well as under ambient conditions reveal a clearly enhanced electromagnetic interaction between the magnetic tip and the nanodots. The dissipated energy per cycle is comparable to the value found for metals, indicating that the interaction of the ion with the target material leads to the creation of metallic Ca nanodots on the surface

Prognostic impact of additional HPV diagnostics in 102 patients with p16-stratified advanced oropharyngeal squamous cell carcinoma

Purpose!#!p16 overexpression was considered as surrogate marker to identify human papillomavirus (HPV)-associated oropharyngeal squamous cell carcinoma (OPSCCs).!##!Methods!#!102 patients with advanced stage OPSCCs treated primarily by transoral lasermicrosurgery were included. Prognostic associations of p16- and HPV-status were analyzed separately and combined.!##!Results!#!In contrast to p16, the HPV-status resulted in no significant survival discrepancies (5-year overall survival (OS) HPV-positive 64.9%, HPV-negative 78.7%). Combining both markers, p16-positive (p16-positive/HPV-positive, p16-positive/HPV-negative) and p16-negative/HPV-negative groups demonstrated comparable high survival (OS 78.1% vs. 85.6% vs. 73.6%). Lowest survival was observed for patients with p16-negative/HPV-positive OPSCCs (OS 40.8%). Never smoking patients with p16-positive OPSCCs demonstrated the highest survival, whereas within former/current smokers with p16-positive and p16-negative disease it was comparable low (OS 90.0% vs. 63.0% vs. 57.4%).!##!Conclusions!#!p16- and HPV-status should not be considered as equivalent markers for a better prognosis. Furthermore, they should not generally predominate patient associated factors like smoking

Ions modulate stress-induced nano-texture in supported fluid lipid bilayers.

Most plasma membranes comprise a large number of different molecules including lipids and proteins. In the standard fluid mosaic model, the membrane function is effected by proteins whereas lipids are largely passive and serve solely in the membrane cohesion. Here we show, using supported 1,2-dioleoyl-sn-glycero-3-phosphocholine lipid bilayers in different saline solutions, that ions can locally induce ordering of the lipid molecules within the otherwise fluid bilayer when the latter is supported. This nanoordering exhibits a characteristic length scale of ∼20 nm, and manifests itself clearly when mechanical stress is applied to the membrane. Atomic force microscopy (AFM) measurements in aqueous solutions containing NaCl, KCl, CaCl2, and Tris buffer show that the magnitude of the effect is strongly ion-specific, with Ca2+ and Tris, respectively, promoting and reducing stress-induced nanotexturing of the membrane. The AFM results are complemented by fluorescence recovery after photobleaching experiments, which reveal an inverse correlation between the tendency for molecular nanoordering and the diffusion coefficient within the bilayer. Control AFM experiments on other lipids and at different temperatures support the hypothesis that the nanotexturing is induced by reversible, localized gel-like solidification of the membrane. These results suggest that supported fluid phospholipid bilayers are not homogenous at the nanoscale, but specific ions are able to locally alter molecular organization and mobility, and spatially modulate the membrane’s properties on a length scale of ∼20 nm. To illustrate this point, AFM was used to follow the adsorption of the membrane-penetrating antimicrobial peptide Temporin L in different solutions. The results confirm that the peptides do not absorb randomly, but follow the ion-induced spatial modulation of the membrane. Our results suggest that ionic effects have a significant impact for passively modulating the local properties of biological membranes, when in contact with a support such as the cytoskeleton

Mechanical heterogeneity of dentin at different length scales as determined by AFM phase contrast

In this study we sought to gain insights of the structural and mechanical heterogeneity of dentin at different length scales. We compared four distinct demineralization protocols with respect to their ability to expose the periodic pattern of dentin collagen. Additionally, we analyzed the phase contrast resulting from AFM images obtained in tapping mode to interrogate the viscoelastic behavior and surface adhesion properties of peritubular and intertubular dentin, and partially demineralized dentin collagen fibrils, particularly with respect to their gap and overlap regions. Results demonstrated that all demineralization protocols exposed the gap and overlap zones of dentin collagen fibrils. Phase contrast analyses suggested that the intertubular dentin, where the organic matrix is concentrated, generated a higher phase contrast due a higher contribution of energy dissipation (damping) than the highly mineralized peritubular region. At increasing amplitudes, viscoelasticity appeared to play a more significant contribution to the phase contrast of the images of collagen fibrils. The overlap region yielded a greater phase contrast than the more elastic gap zones. In summary, our results contribute to the perspective that, at different length scales, dentin is constituted of structural features that retain heterogeneous mechanical properties contributing to overall mechanical performance of the tissue. Furthermore, the interpretation of phase contrast from images generated with AFM tapping mode appears to be an effective tool to gain an improved understanding of the structure and property relationship of biological tissues and biomaterials at the micro- and nano-scale

Skeletal Mineralization Deficits and Impaired Biogenesis and Function of Chondrocyte-Derived Matrix Vesicles in Phospho1(-/-) and Phospho1/Pit1 Double Knockout Mice

International audienceWe have previously shown that ablation of either the Phospho1 or Alpl gene, encoding PHOSPHO1 and tissue-nonspecific alkaline phosphatase (TNAP) respectively, lead to hyperosteoidosis, but that their chondrocyte-derived and osteoblast-derived matrix vesicles (MVs) are able to initiate mineralization. In contrast, the double ablation of Phospho1 and Alpl completely abolish initiation and progression of skeletal mineralization. We argued that MVs initiate mineralization by a dual mechanism: PHOSPHO1-mediated intravesicular generation of inorganic phosphate (Pi ) and phosphate transporter-mediated influx of Pi . To test this hypothesis, we generated mice with col2a1-driven Cre-mediated ablation of Slc20a1, hereafter referred to as Pi t1, alone or in combination with a Phospho1 gene deletion. Pi t1(col2/col2) mice did not show any major phenotypic abnormalities, whereas severe skeletal deformities were observed in the [Phospho1(-/-) ; Pi t1(col2/col2) ] double knockout mice that were more pronounced than those observed in the Phospho1(-/-) mice. Histological analysis of [Phospho1(-/-) ; Pi t1(col2/col2) ] bones showed growth plate abnormalities with a shorter hypertrophic chondrocyte zone and extensive hyperosteoidosis. The [Phospho1(-/-) ; Pi t1(col2/col2) ] skeleton displayed significant decreases in BV/TV%, trabecular number, and bone mineral density, as well as decreased stiffness, decreased strength, and increased postyield deflection compared to Phospho1(-/-) mice. Using atomic force microscopy we found that ∼80% of [Phospho1(-/-) ; Pi t1(col2/col2) ] MVs were devoid of mineral in comparison to ∼50% for the Phospho1(-/-) MVs and ∼25% for the WT and Pi t1(col2/col2) MVs. We also found a significant decrease in the number of MVs produced by both Phospho1(-/-) and [Phospho1(-/-) ; Pi t1(col2/col2) ] chondrocytes. These data support the involvement of phosphate transporter 1, hereafter referred to as Pi T-1, in the initiation of skeletal mineralization and provide compelling evidence that PHOSPHO1 function is involved in MV biogenesis. © 2016 American Society for Bone and Mineral Research

Advances in atomic force microscopy

This article reviews the progress of atomic force microscopy (AFM) in ultra-high vacuum, starting with its invention and covering most of the recent developments. Today, dynamic force microscopy allows to image surfaces of conductors \emph{and} insulators in vacuum with atomic resolution. The mostly used technique for atomic resolution AFM in vacuum is frequency modulation AFM (FM-AFM). This technique, as well as other dynamic AFM methods, are explained in detail in this article. In the last few years many groups have expanded the empirical knowledge and deepened the theoretical understanding of FM-AFM. Consequently, the spatial resolution and ease of use have been increased dramatically. Vacuum AFM opens up new classes of experiments, ranging from imaging of insulators with true atomic resolution to the measurement of forces between individual atoms.Comment: In press (Reviews of Modern Physics, scheduled for July 2003), 86 pages, 44 figure