Stanislav V. Verkhoturov

Nanoscopic Cylindrical Dual Concentric and Lengthwise Block Brush Terpolymers as Covalent Preassembled High-Resolution and High-Sensitivity Negative-Tone Photoresist Materials

We describe a high-resolution, high-sensitivity negative-tone photoresist technique that relies on bottom-up preassembly of differential polymer components within cylindrical polymer brush architectures that are designed to align vertically on a substrate and allow for top-down single-molecule line-width imaging. By applying cylindrical diblock brush terpolymers (DBTs) with a high degree of control over the synthetic chemistry, we achieved large areas of vertical alignment of the polymers within thin films without the need for supramolecular assembly processes, as required for linear block copolymer lithography. The specially designed chemical compositions and tuned concentric and lengthwise dimensions of the DBTs enabled high-sensitivity electron-beam lithography of patterns with widths of only a few DBTs (sub-30 nm line-width resolution). The high sensitivity of the brush polymer resists further facilitated the generation of latent images without postexposure baking, providing a practical approach for controlling acid reaction/diffusion processes in photolithography.

Micropatterning of Proteins and Mammalian Cells on Indium Tin Oxide

This paper describes a novel surface engineering approach that combines oxygen plasma treatment and electrochemical activation to create micropatterned cocultures on indium tin oxide (ITO) substrates. In this approach, photoresist was patterned onto an ITO substrate modified with poly(ethylene) glycol (PEG) silane. The photoresist served as a stencil during exposure of the surface to oxygen plasma. Upon incubation with collagen (I) solution and removal of the photoresist, the ITO substrate contained collagen regions surrounded by nonfouling PEG silane. Chemical analysis carried out with time-of-flight secondary ion mass spectrometry (ToF-SIMS) at different stages in micropatterned construction verified removal of PEG-silane during oxygen plasma and presence of collagen and PEG molecules on the same surface. Imaging ellipsometry and atomic force microscopy (AFM) were employed to further investigate micropatterned ITO surfaces. Biological application of this micropatterning strategy was demonstrated through selective attachment of mammalian cells on the ITO substrate. Importantly, after seeding the first cell type, the ITO surfaces could be activated by applying negative voltage (-1.4 V vs Ag/AgCl). This resulted in removal of nonfouling PEG layer and allowed to attach another cell type onto the same surface and to create micropatterned cocultures. Micropatterned cocultures of primary hepatocytes and fibroblasts created by this strategy remained functional after 9 days as verified by analysis of hepatic albumin. The novel surface engineering strategy described here may be used to pattern multiple cell types on an optically transparent and conductive substrate and is envisioned to have applications in tissue engineering and biosensing.

A comparative study of the cytotoxicity and corrosion resistance of nickel-titanium and titanium-niobium shape memory alloys.

Nickel-titanium (NiTi) shape memory alloys (SMAs) are commonly used in a range of biomedical applications. However, concerns exist regarding their use in certain biomedical scenarios due to the known toxicity of Ni and conflicting reports of NiTi corrosion resistance, particularly under dynamic loading. Titanium-niobium (TiNb) SMAs have recently been proposed as an alternative to NiTi SMAs due to the biocompatibility of both constituents, the ability of both Ti and Nb to form protective surface oxides, and their superior workability. However, several properties critical to the use of TiNb SMAs in biomedical applications have not been systematically explored in comparison with NiTi SMAs. These properties include cytocompatibility, corrosion resistance, and alterations in alloy surface composition in response to prolonged exposure to physiological solutions. Therefore, the goal of the present work was to comparatively investigate these aspects of NiTi (49.2 at.% Ti) and TiNb (26 at.% Nb) SMAs. The results from the current studies indicate that TiNb SMAs are less cytotoxic than NiTi SMAs, at least under static culture conditions. This increased TiNb cytocompatibility was correlated with reduced ion release as well as with increased corrosion resistance according to potentio-dynamic tests. Measurements of the surface composition of samples exposed to cell culture medium further supported the reduced ion release observed from TiNb relative to NiTi SMAs. Alloy composition depth profiles also suggested the formation of calcium phosphate deposits within the surface oxide layers of medium-exposed NiTi but not of TiNb. Collectively, the present results indicate that TiNb SMAs may be promising alternatives to NiTi for certain biomedical applications.

Analysis of Native Biological Surfaces Using a 100 kV Massive Gold Cluster Source

In the present work, the advantages of a new, 100 kV platform equipped with a massive gold cluster source for the analysis of native biological surfaces are shown. Inspection of the molecular ion emission as a function of projectile size demonstrates a secondary ion yield increase of ~100× for 520 keV Au(400)(4+) as compared to 130 keV Au(3)(1+) and 43 keV C(60). In particular, yields of tens of percent of molecular ions per projectile impact for the most abundant components can be observed with the 520 keV Au(400)(4+) probe. A comparison between 520 keV Au(400)(4+) time-of-flight-secondary ion mass spectrometry (TOF-SIMS) and matrix assisted laser desorption ionization-mass spectrometry (MALDI-MS) data showed a similar pattern and similar relative intensities of lipid components across a rat brain sagittal section. The abundant secondary ion yield of analyte-specific ions makes 520 keV Au(400)(4+) projectiles an attractive probe for submicrometer molecular mapping of native surfaces.

Characterization of individual ag nanoparticles and their chemical environment.

Silver nanoparticles (NPs) of approximately 5 nm diameter deposited in a single layer on glycine were examined with cluster secondary ion mass spectrometry (SIMS) in the event-by-event bombardment-detection mode. The projectiles used were Au(3)(+), C(60)(+), and Au(400)(4+) with impact energies of 34, 26, and 136 keV, respectively. The highest secondary ion yields were obtained with Au(400)(4+). The method presented can test single or multilayer organization and determine surface coverage. NPs are identified one-by-one for chemical composition. Chemical and physical information in a nonimaging mode was resolved at approximately 10 nm. Grazing vs direct impacts on NPs were identified and quantified. NP fragmentation from direct impacts with Au(400)(4+) and C(60)(+) was observed for the first time.

Molecular identification of individual nano-objects.

Secondary ion mass spectrometry (SIMS) run in the event-by-event bombardment/detection mode provides a unique ability to obtain molecular information from single nano-objects, since assays are based on secondary ion coemission from single impacts. The characterization of individual nano-objects is demonstrated with negatively charged polymer spheres that are attracted to and retained by nanoalumina whiskers. The whiskers, 2 nm in diameter and approximately 250 nm in length, are grafted to a microglass fiber with an average diameter of approximately 0.6 microm and several millimeters long. The spheres are monodisperse polystyrene nanoparticles (30 nm diameter). Massive Au projectiles, specifically 136 keV Au(400)(4+), were utilized to bombard analyte surfaces due to its high efficiency for producing multi-ion emission identified by time-of-flight mass spectrometry. Our results show that this mode of mass spectrometry can provide information on the nature, size, relative location, and abundance of nano-objects in the field of view. The key to characterizing nanodomains is to monitor the coincidental secondary ion emission from the nanovolume perturbed by single projectile impacts.

Characterization of individual nano-objects by secondary ion mass spectrometry.

We present secondary ion mass spectrometry (SIMS) data obtained from the bombardment of a novel nanomaterial with a suite of projectiles: Au1+, Au3+, Au9+, and Au400(4+). These are the first experiments where free-standing nano-objects were bombarded with kiloelectronvolt projectiles of atomic to nanoparticle size (Au400(4+)). The objects are aluminum monohydrate nanowhiskers, identified as crystalline boehmite (AlOOH) using X-ray diffraction. The nanoalumina is bonded to a microglass fiber that serves as a scaffold. The largest projectile, Au400(4+), has a diameter of approximately 2 nm, comparable to the nominal diameter of the nanowhiskers. There are notable differences in secondary ion (SI) response from sample volumes too small for full projectile energy deposition. The whisker spectra are dominated by small clusters--the most abundant species being AlO- and AlO2-. Bulk samples have larger yields for AlO2- than AlO-, whereas this trend is reversed in the whisker samples. Bulk samples give similar abundances of large SI cluster families [(Al2O3)(n)AlO2]- and [(Al2O3)(n)OH]-, whereas the whisker samples give an order of magnitude lower yield of these SIs. Given the nature of our experiments, i.e., the event-by-event bombardment/detection mode, we are uniquely able to obtain information from SIs emitted from single-projectile impacts. As such, effective yields were calculated in order to determine quantitative differences between the nano-objects and bulk samples.

Electrochemical and Structural Effects of In Situ Li2O Extraction from Li2MnO3 for Li-Ion Batteries

Li2MnO3 is an attractive cathode material due to its low cost, nontoxicity and potentially high capacity. However, its electrochemical inactivity, its poor electronic conductivity, and uncertainty about its underlying mechanism have limited its development. In this work, an in situ technique for extraction of Li and O during deposition of the thin film cathode is developed to investigate structural and electrochemical effects in a controlled fashion. MnO2 has been observed in samples with severe O and Li deficiency (capacity of 115 mAh g(-1)), while Li2MnO3 cathodes with slight excess of O and Li (capacity of 225 mAh g(-1)) can be synthesized by tuning growth conditions appropriately. Formation of a MnO2 phase, especially in Li and O deficient structures, could be a possible reason for irreversible capacity loss in Li2MnO3 related materials. Further investigation into stoichiometric and microstructure variations enabled by this technique allows rapid investigation of Li2MnO3 as well as other Li-rich composites.

Examination of nanoparticles via single large cluster impacts.

This study deals with the determination of the relative abundance of the oxide layer in the near-surface volume of aluminum nanoparticles of 50-100 nm in diameter. They are bombarded with a sequence of single projectiles of Au 400(4+) accelerated to 136 keV. The ionized ejecta from each impact are recorded individually which allows identification of ions emitted from a surface volume of approximately 10 nm in diameter and 5-10 nm in depth. The mode of analyzing ejecta individually from each single cluster impact is a means to apply mass spectrometry in nanovolumes.

Preventing adhesion of Escherichia coli O157:H7 and Salmonella Typhimurium LT2 on tomato surfaces via ultrathin polyethylene glycol film.

This work deals with adhesion of Escherichia coli O157:H7 and Salmonella enterica subsp. enterica serovar Typhimurium LT2 (S. Typhimurium LT2) on polyethylene glycol (PEG) coated tomato surfaces. PEG coating was characterized by water contact angle technique, scanning electron microscopy, and secondary ion mass spectrometry. It was shown that PEG films could physisorb on the tomato surfaces after the oxygen plasma treatment, which made some outermost layers of the surfaces hydrophilic. Bacterial adhesion on PEG coated tomato surface was studied by standard plate count, fluorescence microscopy, and scanning electron microscopy techniques. Fully covered PEG film reduced the bacterial attachment 90% or more in comparison to the bare tomato surface. The degree of bacterial attachment decreased exponentially with increasing PEG coverage. When desired, PEG film could be removed by rinsing with water. Overall, this work demonstrates the proof-of-concept that an ultrathin film of polyethylene glycol may be used to effectively inhibit the attachment of pathogenic bacteria on tomato surfaces.