Peter Manfred Schön

Interlaboratory round robin on cantilever calibration for AFM force spectroscopy

Single-molecule force spectroscopy studies performed by Atomic Force Microscopes (AFMs) strongly rely on accurately determined cantilever spring constants. Hence, to calibrate cantilevers, a reliable calibration protocol is essential. Although the thermal noise method and the direct Sader method are frequently used for cantilever calibration, there is no consensus on the optimal calibration of soft and V-shaped cantilevers, especially those used in force spectroscopy. Therefore, in this study we aimed at establishing a commonly accepted approach to accurately calibrate compliant and V-shaped cantilevers. In a round robin experiment involving eight different laboratories we compared the thermal noise and the Sader method on ten commercial and custom-built AFMs. We found that spring constants of both rectangular and V-shaped cantilevers can accurately be determined with both methods, although the Sader method proved to be superior. Furthermore, we observed that simultaneous application of both methods on an AFM proved an accurate consistency check of the instrument and thus provides optimal and highly reproducible calibration. To illustrate the importance of optimal calibration, we show that for biological force spectroscopy studies, an erroneously calibrated cantilever can significantly affect the derived (bio)physical parameters. Taken together, our findings demonstrated that with the pre-established protocol described reliable spring constants can be obtained for different types of cantilevers.

The effect of PeakForce tapping mode AFM imaging on the apparent shape of surface nanobubbles

Until now, TM AFM (tapping mode or intermittent contact mode atomic force microscopy) has been the most often applied direct imaging technique to analyze surface nanobubbles at the solid-aqueous interface. While the presence and number density of nanobubbles can be unequivocally detected and estimated, it remains unclear how much the a priori invasive nature of AFM affects the apparent shapes and dimensions of the nanobubbles. To be able to successfully address the unsolved questions in this field, the accurate knowledge of the nanobubbles dimensions, radii of curvature etc is necessary. In this contribution we present a comparative study of surface nanobubbles on HOPG (highly oriented pyrolytic graphite) in water acquired with (i) TM AFM and (ii) the recently introduced PFT (PeakForce tapping) mode, in which the force exerted on the nanobubbles rather than the amplitude of the resonating cantilever is used as the AFM feedback parameter during imaging. In particular, we analyzed how the apparent size and shape of nanobubbles depend on the maximum applied force in PFT AFM. Even for forces as small as 73 pN, the nanobubbles appeared smaller than their true size, which was estimated from an extrapolation of the bubble height to zero applied force. In addition, the size underestimation was found to be more pronounced for larger bubbles. The extrapolated true nanoscopic contact angles for nanobubbles on HOPG, measured in PFT AFM, ranged from 145°xa0to 175°xa0and were only slightly underestimated by scanning with non-zero forces. This result was comparable to the nanoscopic contact angles of 160°-175°xa0measured using TM AFM in the same set of experiments. Both values disagree, in accordance with the literature, with the macroscopic contact angle of water on HOPG, measured here to be 63°xa0±xa02°.

Characterization and molecular engineering of surface-grafted polymer brushes across the length scales by atomic force microscopy

With the advent of regulated, surface initiated polymerizations, specifically using controlled radical approaches, the choice of polymerizable compounds and the control over grafting chemistry have seen tremendous advancement. New analysis techniques and approaches are now needed to characterize these brushes with molecular precision. In addition, spatial structure control at the nanoscale, and tuning of thickness as well as composition of the brushes, have become feasible by utilizing recently developed enabling molecular nanofabrication approaches. Atomic force microscopy (AFM) is a powerful analytical tool for the characterization of polymer brushes, as well as for the fabrication of brush structures across the length scales. AFM has been used to investigate polymer brushes in a number of ways including imaging surface morphologies, measuring brush thickness, estimating the value of number average molar mass, observing stimulus responsive behavior and probing surface mechanical properties. In addition, AFM based methods such as nanoscratching, dip-pen nanolithography (DPN) and scanning probe oxidation (SPO) have been also employed for the nanofabrication of patterned polymer brushes. This feature article gives a short account of this field and highlights recent advances.

Preparation and Characterization of Multilayer Coated Microdroplets : Droplet Deformation Simultaneously Probed by Atomic Force Spectroscopy and Optical Detection

We report the preparation and characterization of multilayer coated droplets in an emulsion. Stability and control of mass transport across the interface are the key issues for such coated microdroplets. Shelf life of cosmetic, pharmaceutical, and food formulations can be improved by increasing stability. Moreover, such emulsions have potential applications in drug delivery and storage. A primary oil-in-water emulsion with caseinate as an emulsifier was prepared. On the basis of attractive electrostatic interactions, polyelectrolytes with opposite charges were added layer by layer. The oil droplets (particle size around 10 microm) were successively coated with casein, pectin, whey proteins, pectin, whey proteins, and pectin. Laser diffraction spectroscopy, particle charge measurements, and confocal laser scanning microscopy were applied to characterize the multilayer droplets. The complementary results indicate that the inner layers merge and the packing density of the interface increases. AFM-induced mechanical compression of single oil droplets coated with casein and pectin is monitored by an inverted optical microscope, and simultaneously AFM force curves are recorded. Thus, the deformation of the droplet is reflected by its lateral expansion and the force curve. Force volume imaging is applied to probe the lateral distribution of mechanical properties of the droplet.

Sieving of Hot Gases by Hyper-Cross-Linked Nanoscale-Hybrid Membranes

Macromolecular networks consisting of homogeneously distributed covalently bonded inorganic and organic precursors are anticipated to show remarkable characteristics, distinct from those of the individual constituents. A novel hyper-cross-linked ultrathin membrane is presented, consisting of a giant molecular network of alternating polyhedral oligomeric silsesquioxanes and aromatic imide bridges. The hybrid characteristics of the membrane are manifested in excellent gas separation performance at elevated temperatures, providing a new and key enabling technology for many important industrial scale applications.

Dynamics of Decanethiol Self-Assembled Monolayers on Au(111) Studied by Time-Resolved Scanning Tunneling Microscopy

We investigated the dynamics of decanethiol self-assembled monolayers on Au(111) surfaces using time-resolved scanning tunneling microscopy at room temperature. The expected ordered phases (β, δ, χ*, and φ) and a disordered phase (ε) were observed. Current-time traces with the feedback loop disabled were recorded at different locations on the surface. The sulfur end group of the decanethiolate molecule exhibits a stochastic two-level switching process when the molecule is adsorbed in a (local) β phase registry. This two-level process is attributed to the diffusion of the Au-thiolate complex between two adjacent adsorption sites. The irregular current jumps in the current-time traces recorded on the tails of decanethiolate molecules in the ordered β, δ, and χ* phases are ascribed to wagging of the alkyl tails. Finally, the disordered phase is characterized by even larger current jumps, which indicates that the tail of the decanethiolate flips up occasionally and makes contact with the tip. Our experiments reveal that the massive dynamics of the self-assembled monolayer is due to diffusion of decanethiol-Au complexes, rather than the diffusion of decanethiolate molecules.

Probing biofouling resistant polymer brush surfaces by atomic force microscopy based force spectroscopy

The protein repellency and biofouling resistance of zwitterionic poly(sulfobetaine methacrylate)(pSBMA) brushes grafted via surface initiated polymerization (SIP) from silicon and glass substrata was assessed using atomic force microscopy (AFM) adherence experiments. Laboratory settlement assays were conducted with cypris larvae of the barnacle Balanus amphitrite. AFM adherence includes the determination of contact rupture forces when AFM probe tips are withdrawn from the substratum. When the surface of the AFM tip is modified, adherence can be assessed with chemical specifity using a method known as chemical force microscopy (CFM). In this study, AFM tips were chemically functionalized with (a) fibronectin- here used as model for a nonspecifically adhering protein - and (b) arginine-glycine-aspartic acid (RGD) peptide motifs covalently attached to poly(methacrylic acid) (PMAA) brushes as biomimics of cellular adhesion receptors. Fibronectin functionalized tips showed significantly reduced nonspecific adhesion to pSBMA-modified substrata compared to bare gold (2.3±0.75 nN) and octadecanethiol (ODT) self-assembled monolayers (1.3±0.75 nN). PMAA and PMAA-RGD modified probes showed no significant adhesion to pSBMA modified silicon substrata. The results gathered through AFM protein adherence studies were complemented by laboratory fouling studies, which showed no adhesion of cypris larvae of Balanus amphitrite on pSBMA. With regard to its unusually high non-specific adsorption to a wide variety of materials the behavior of fibronectin is analogous to the barnacle cyprid temporary adhesive that also binds well to surfaces differing in polarity, charge and free energy. The antifouling efficacy of pSBMA may, therefore, be directly related to the ability of this surface to resist nonspecific protein adsorption.

Molecular dynamics and energy landscape of decanethiolates in self-assembled monolayers on Au(111) by STM

The energetics and dynamics of the various phases of decanethiolate self-assembled monolayers on Au(111) surfaces were studied with scanning tunneling microscopy. We have observed five different phases of the decanethiolate monolayer on Au(111): four ordered phases (β, δ, χ*, and φ) and one disordered phase (ε). We have determined the boundary free energies between the disordered and order phases by analyzing the thermally induced meandering of the domain boundaries. On the basis of these results, we are able to accurately predict the two-dimensional phase diagram of the decanethiolate/Au(111) system. The order-disorder phase transition of the χ* phase occurs at 295 K, followed by the order-disorder phase transition of the β phase at 325 K. Above temperatures of 325 K, only the densely packed φ and disordered ε phases remain. Our findings are in good agreement with the phase diagram of the decanethiolate/Au(111) system that was put forward by Poirier et al. [Langmuir 2001, 17 (4), 1176-1183].

Imaging and force probing RNA by atomic force microscopy.

In the past 30years, the atomic force microscope (AFM) has become a true enabling platform in the life sciences opening entire novel avenues for structural and dynamic studies of biological systems. It enables visualization, probing and manipulation across the length scales, from single molecules to living cells in buffer solution under physiological conditions without the need for labeling or staining of the specimen. In particular, for structural studies of nucleic acids and assemblies thereof, the AFM has matured into a routinely used tool providing nanometer spatial resolution. This includes ssRNA, dsRNA and nucleoprotein complexes thereof, as well as RNA aggregates and 2D RNA assemblies. By AFM unique information can be obtained on RNA based assemblies which are becoming increasingly important as novel unique building blocks in the emerging field of RNA nanotechnology. In addition, the AFM is of fundamental relevance to study biological relevant RNA interactions and dynamics. In this short review first the basic functioning principles of commonly used AFM modes including AFM based force spectroscopy will be briefly described. Next a brief overview will be given on structural studies that have been done related to AFM topographic imaging of RNA, RNA assemblies and aggregates. Finally, an overview on AFM beyond imaging will be provided. This includes force spectroscopy of RNA under physiological conditions in aqueous buffer to probe RNA interaction with proteins and ligands as well as other AFM tip based RNA probing. The main intention of this short review to give the reader a flavor of what AFM contributes to RNA research and engineering.

Austenitic and duplex stainless steels in simulated physiological solution characterized by electrochemical and X-ray photoelectron spectroscopy studies.

A study of oxide layers grown on 2205 duplex stainless steel (DSS) and AISI 316L austenitic stainless steel in simulated physiological solution is presented here in order to establish the possibility of replacement of AISI 316 L with 2205 DSS in biomedical applications. The results of the potentiodynamic measurements show that the extent of the passive range significantly increased for DSS 2205 compared to AISI 316L stainless steel. Cyclic voltammetry was used to investigate electrochemical processes taking place on the steel surfaces. Oxide layers formed by electrochemical oxidation at different oxidation potentials were studied by X-ray photoelectron spectroscopy, and their compositions were analyzed as a function of depth. The main constituents on both the investigated materials were Cr- and Fe-oxides. Atomic force microscopy topography studies revealed the higher corrosion resistance of the DSS 2205 compared to the AISI 316L under the chosen experimental conditions.