Removal of electrostatic artifacts in magnetic force microscopy by controlled magnetization of the tip: application to superparamagnetic nanoparticles

Magnetic force microscopy (MFM) has been demonstrated as valuable technique for the characterization of magnetic nanomaterials. To be analyzed by MFM techniques, nanomaterials are generally deposited on flat substrates, resulting in an additional contrast in MFM images due to unavoidable heterogeneous electrostatic tip-sample interactions, which cannot be easily distinguished from the magnetic one. In order to correctly interpret MFM data, a method to remove the electrostatic contributions from MFM images is needed. In this work, we propose a new MFM technique, called controlled magnetization MFM (CM-MFM), based on the in situ control of the probe magnetization state, which allows the evaluation and the elimination of electrostatic contribution in MFM images. The effectiveness of the technique is demonstrated through a challenging case study, i.e., the analysis of superparamagnetic nanoparticles in absence of applied external magnetic field. Our CM-MFM technique allowed us to acquire magnetic images depurated of the electrostatic contributions, which revealed that the magnetic field generated by the tip is sufficient to completely orient the superparamagnetic nanoparticles and that the magnetic tip-sample interaction is describable through simple models once the electrostatic artifacts are removed

Self-assembling of calcium salt of the new DNA base 5-carboxylcytosine

Supramolecular architectures involving DNA bases can have a strong impact in several fields such as nanomedicine and nanodevice manufacturing. To date, in addition to the four canonical nucleobases (adenine, thymine, guanine and cytosine), four other forms of cytosine modified at the 5 position have been identified in DNA. Among these four new cytosine derivatives, 5-carboxylcytosine has been recently discovered in mammalian stem cell DNA, and proposed as the final product of the oxidative epigenetic demethylation pathway on the 5 position of cytosine. In this work, a calcium salt of 5-carboxylcytosine has been synthesized and deposited on graphite surface, where it forms self-assembled features as long range monolayers and up to one micron long filaments. These structures have been analyzed in details combining different theoretical and experimental approaches: X-ray single-crystal diffraction data were used to simulate the molecule-graphite interaction, first using molecular dynamics and then refining the results using density functional theory (DFT); finally, data obtained with DFT were used to rationalize atomic force microscopy (AFM) results

Detection of stiff nanoparticles within cellular structures by contact resonance atomic force microscopy subsurface nanomechanical imaging

Detecting stiff nanoparticles buried in soft biological matrices by atomic force microscopy (AFM) based techniques represents a new frontier in the field of scanning probe microscopies, originally developed as surface characterization methods. Here we report the detection of stiff (magnetic) nanoparticles (NPs) internalized in cells by using contact resonance AFM (CR-AFM) employed as a potentially non-destructive subsurface characterization tool. Magnetite (Fe3O4) NPs were internalized in microglial cells from cerebral cortices of mouse embryos of 18 days by phagocytosis. Nanomechanical imaging of cells was performed by detecting the contact resonance frequencies (CRFs) of an AFM cantilever held in contact with the sample. Agglomerates of NPs internalized in cells were visualized on the basis of the local increase in the contact stiffness with respect to the surrounding biological matrix. A second AFM-based technique for nanomechanical imaging, i.e., HarmoniX™, as well as magnetic force microscopy and light microscopy were used to confirm the CR-AFM results. Thus, CR-AFM was emonstrated as a promising technique for subsurface imaging of nanomaterials in biological samples

Synthèse et caractérisation multiéchelle de matériaux et de systèmes pour applications biomédicales

A procedure aimed at designing innovative epoxy resin-free sandwich materials (i.e., layered structure composed of two metal skin and a polymer core) able to reduce stress-shielding effect at the implant/bone interface was developed. For this purpose, titanium (Ti) and poly methymethacrilate (PMMA), the most extensively materials used for biomedical applications, were employed. In particular, surface-confined PMMA layers were proposed as adhesives to stick a PMMA foil (used as core of the structure) on the metallic Ti skin sheets exploiting the miscibility between the tethered polymer chains (previously grown on the Ti) and those of an adhering PMMA foil.To this purpose, a three steps strategy based on a suitable functionalization of Ti surface was developed. First of all, a chemical activation of Ti surface was performed. Then, a “grafting from” method was used to immobilize a polymerization initiator on the activated Ti surface. Finally, the polymer chains were grown from the initiator-modified surfaces using a surface initiation atom transfer radical polymerization (SI-ATRP). Biocompatible Ti/PMMA/Ti sandwiches were then prepared by hot-pressing, inserting between the two PMMA-coated Ti surfaces a thick PMMA foil.Des matériaux sandwichs fabriqués sans colle époxy on été conçu pour réduire les contraintes mécaniques, ou “stress shielding”, entre l’os environnant et l’implant. Le titane (Ti) et le polyméthacrylate de méthyle (PMMA) sont les matériaux les plus utilisés dans les applications biomédicales, et on été choisi comme composants de base. Pour cela, on a élaboré des interfaces Ti/polymère dans lesquelles le métal et le polymère sont liés par une liaison covalente; cette couche de polymère permettra ultérieurement l’adhésion entre le métal et une feuille de polymère qui constituera le cœur du sandwich. Dans ce but, une stratégie en trois étapes permettant d’obtenir une fonctionnalisation de la surface du titane a été développé. Tout d’abord, la surface du Ti a été activée chimiquement; ensuite un initiateur de polymérisation y a été greffé de façon covalente. Enfin, la croissance des chaines polymères a été obtenue en utilisant une polymérisation par transfert d’atomes à partir de l’initiateur (SI-ATRP). Les sandwichs ont été préparés en insérant une feuille de polymère entre les deux feuilles de Ti recouvertes de polymère greffé et en pressant les trois composants à une température supérieure à celle de la transition vitreuse du polymère

Elastic modulus measurements at variable temperature: Validation of atomic force microscopy techniques

The development of polymer-based nanocomposites to be used in critical thermal environments requires the characterization of their mechanical properties, which are related to their chemical composition, size, morphology and operating temperature. Atomic force microscopy (AFM) has been proven to be a useful tool to develop techniques for the mechanical characterization of these materials, thanks to its nanometer lateral resolution and to the capability of exerting ultra-low loads, down to the piconewton range. In this work, we demonstrate two techniques, one quasi-static, i.e., AFM- based indentation (I-AFM), and one dynamic, i.e., contact resonance AFM (CR-AFM), for the mechanical characterization of compliant materials at variable temperature. A cross-validation of I-AFM and CR-AFM has been performed by comparing the results obtained on two reference materials, i.e., low-density polyethylene (LDPE) and polycarbonate (PC), which demonstrated the accuracy of the techniques

Magnetic Force Microscopy

Magnetic force microscopy (MFM) refers to a family of scanning probe techniques based on atomic force microscopy (AFM), which allow one to image the magnetic properties of the sample surface at the nanoscale, simultaneously to its topography. Here, we review the most widespread MFM techniques, mainly dynamic MFM although static MFM is also briefly described for the sake of completeness. We illustrate the working principles, the experimental setups, and the analytical models describing the MFM response, which are fundamental for understanding and quantitatively interpreting the contrast in MFM images. An overview is given of the application fields of MFM, which cover almost all the magnetic materials, from recording media to ferromagnetic materials, nanomaterials and nanoparticles, alone and in organic or biological systems. Finally, some advances, hot topics, new applications, and open issues are presented, including the effect of external magnetic fields, nonmagnetic interactions, MFM tips calibration and advanced probes, and magnetic imaging with variable temperature

Identification of nanoparticles and nanosystems in biological matrices with scanning probe microscopy

Identification of nanoparticles and nanosystems into cells and biological matrices is a hot research topic in nanobiotechnologies. Because of their capability to map physical properties (mechanical, electric, magnetic, chemical, or optical), several scanning probe microscopy based techniques have been proposed for the subsurface detection of nanomaterials in biological systems. In particular, atomic force microscopy (AFM) can be used to reveal stiff nanoparticles in cells and other soft biomaterials by probing the sample mechanical properties through the acquisition of local indentation curves or through the combination of ultrasound-based methods, like contact resonance AFM (CR-AFM) or scanning near field ultrasound holography. Magnetic force microscopy can detect magnetic nanoparticles and other magnetic (bio)materials in nonmagnetic biological samples, while electric force microscopy, conductive AFM, and Kelvin probe force microscopy can reveal buried nanomaterials on the basis of the differences between their electric properties and those of the surrounding matrices. Finally, scanning near field optical microscopy and tip-enhanced Raman spectroscopy can visualize buried nanostructures on the basis of their optical and chemical properties. Despite at a still early stage, these methods are promising for detection of nanomaterials in biological systems as they could be truly noninvasive, would not require destructive and time-consuming specific sample preparation, could be performed in vitro, on alive samples and in water or physiological environment, and by continuously imaging the same sample could be used to dynamically monitor the diffusion paths and interaction mechanisms of nanomaterials into cells and biological systems. This article is categorized under: Diagnostic Tools > In Vivo Nanodiagnostics and Imaging Nanotechnology Approaches to Biology > Nanoscale Systems in Biology

Design of Optimized PEDOT‐Based Electrodes for Enhancing Performance of Living Photovoltaics Based on Phototropic Bacteria

Living photovoltaics represent a growing class of microbial devices that are based on whole cell–electrode interactions. The limited charge transfer at the cell–electrode interface represents a significant bottleneck in realizing an efficient technology. This study focuses on the development of poly(3,4‐ethylenedioxythiophene) (PEDOT)‐based electrodes that are electrosynthesized in the presence of a sodium dodecyl sulphate (SDS) dopant. Potentiodynamic and potentiostatic electrochemical techniques, as well as scanning electron microscopy (SEM), atomic force microscopy (AFM), Raman spectroscopy, and theoretical modelling of the electropolymerization transient, are employed to create and characterize PEDOT electrodes under various conditions. The electrodes are able to capture photosynthetically derived current under multiple light–dark cycles when interfaced with Synechocystis sp. PCC 6803. In the presence of the Synechocystis, the PEDOT electrodes show a sixfold and twofold enhancement over conventional graphite electrodes for both mediatorless and K3Fe(CN)6‐mediated conditions, respectively. The ability of these electrodes to enhance extracted photocurrent for both direct and indirect electron transfer mechanisms provides a versatile platform for improving various microbial devices

Tailored extracellular electron transfer pathways enhance the electroactivity of Escherichia coli

Extracellular electron transfer (EET) engineering in Escherichia coli holds great potential for bioremediation, energy and electrosynthesis applications fueled by readily available organic substrates. Due to its vast metabolic capabilities and availability of synthetic biology tools to adapt strains to specific applications, E. coli is of advantage over native exoelectrogens, but limited in electron transfer rates. We enhanced EET in engineered strains through systematic expression of electron transfer pathways differing in cytochrome composition, localization and origin. While a hybrid pathway harboring components of an E. coli nitrate reductase and the Mtr complex from the exoelectrogen Shewanella oneidensis MR-1 enhanced EET, the highest efficiency was achieved by implementing the complete Mtr pathway from S. oneidensis MR1 in E. coli. We show periplasmic electron shuttling through overexpression of a small tetraheme cytochrome to be central to the electroactivity of this strain, leading to enhanced degradation of the pollutant methyl orange and significantly increased electrical current to graphite electrodes.LN

Electrochemical atomic force microscopy: In situ monitoring of electrochemical processes

The in-situ electrodeposition of polyaniline (PANI), one of the most attractive conducting polymers (CP), has been monitored performing electrochemical atomic force microscopy (EC-AFM) experiments. The electropolymerization of PANI on a Pt working electrode has been observed performing cyclic voltammetry experiments and controlling the evolution of current flowing through the electrode surface, together with a standard AFM image. The working principle and the potentialities of this emerging technique are briefly reviewed and factors limiting the studying of the in-situ electrosynthesis of organic compounds discussed