monitoring-conformational-dynamics-single-unmodified-proteins-using

Monitoring Conformational Dynamics of Single Unmodified Proteins using Plasmonic Nanotweezers

Current single-molecule techniques to characterize proteins typically require labels, tethers, or the use of non-native solution conditions. Such changes ...

University of Nottingham

With the rapidly expanding industrial and research applications of near-critical and supercritical technology there is a pressing need for a simple and inexpensive sensor that may be used to determine the phase coexistence regions of fluid mixtures and to establish whether a fluid system is below, at, or above, a critical point. Mechanically vibrating AT-cut quartz plates may be used to determine the product of the fluid density and viscosity of a fluid in which it is immersed, through measurement of the impedance minimum of the electrical equivalent circuit or of the corresponding frequency. The density-viscosity product changes abruptly between fluid phases and rapidly along the isotherm corresponding to the critical temperature, enabling such a plate to act as a sensor of these fluid features. We consider the limitations and linearity of such a sensor and its behavior when a liquid-gas meniscus crosses its surface. We demonstrate for the first time the effective use of an AT-cut quartz sensor in mapping the phase behavior of fluids, using measurements made on carbon dioxide and ethane for calibration and then investigating an ethane-carbon dioxide mixture. The advantages of this experimental approach are that (i) piezoelectric sensors are available for operation up to 1,000 degrees C and at extremely high pressures and (ii) the measurement of the density-viscosity product of supercritical fluids is inherently simpler than traditional techniques for determining phase behavior.

Probing the vapor-liquid phase behaviors of near-critical and supercritical fluids using a shear mode piezoelectric sensor.

High broad-band photoresponsivity of mechanically formed InSe-graphene van der Waals heterostructures is achieved by exploiting the broad-band transparency of graphene, the direct bandgap of InSe, and the favorable band line up of InSe with graphene. The photoresponsivity exceeds that for other van der Waals heterostructures and the spectral response extends from the near-infrared to the visible spectrum.

High broad-band photoresponsivity of mechanically formed InSe-graphene van der Waals heterostructures.

Graphene grown by high temperature molecular beam epitaxy on hexagonal boron nitride (hBN) forms continuous domains with dimensions of order 20 μm, and exhibits moiré patterns with large periodicities, up to ~30 nm, indicating that the layers are highly strained. Topological defects in the moiré patterns are observed and attributed to the relaxation of graphene islands which nucleate at different sites and subsequently coalesce. In addition, cracks are formed leading to strain relaxation, highly anisotropic strain fields, and abrupt boundaries between regions with different moiré periods. These cracks can also be formed by modification of the layers with a local probe resulting in the contraction and physical displacement of graphene layers. The Raman spectra of regions with a large moiré period reveal split and shifted G and 2D peaks confirming the presence of strain. Our work demonstrates a new approach to the growth of epitaxial graphene and a means of generating and modifying strain in graphene.

Strain-Engineered Graphene Grown on Hexagonal Boron Nitride by Molecular Beam Epitaxy.

We demonstrate direct epitaxial growth of high-quality hexagonal boron nitride (hBN) layers on graphite using high-temperature plasma-assisted molecular beam epitaxy. Atomic force microscopy reveals mono- and few-layer island growth, while conducting atomic force microscopy shows that the grown hBN has a resistance which increases exponentially with the number of layers, and has electrical properties comparable to exfoliated hBN. X-ray photoelectron spectroscopy, Raman microscopy and spectroscopic ellipsometry measurements on hBN confirm the formation of sp-bonded hBN and a band gap of 5.9 ± 0.1 eV with no chemical intermixing with graphite. We also observe hexagonal moiré patterns with a period of 15 nm, consistent with the alignment of the hBN lattice and the graphite substrate.

Hexagonal Boron Nitride Tunnel Barriers Grown on Graphite by High Temperature Molecular Beam Epitaxy.

Lattice-matched graphene on hexagonal boron nitride is expected to lead to the formation of a band gap but requires the formation of highly strained material and has not hitherto been realized. We demonstrate that aligned, lattice-matched graphene can be grown by molecular beam epitaxy using substrate temperatures in the range 1600-1710 °C and coexists with a topologically modified moiré pattern with regions of strained graphene which have giant moiré periods up to ∼80 nm. Raman spectra reveal narrow red-shifted peaks due to isotropic strain, while the giant moiré patterns result in complex splitting of Raman peaks due to strain variations across the moiré unit cell. The lattice-matched graphene has a lower conductance than both the Frenkel-Kontorova-type domain walls and also the topological defects where they terminate. We relate these results to theoretical models of band gap formation in graphene/boron nitride heterostructures.

Lattice-Matched Epitaxial Graphene Grown on Boron Nitride.

Monolayer hexagonal boron nitride (hBN) tunnel barriers investigated using conductive atomic force microscopy reveal moiré patterns in the spatial maps of their tunnel conductance consistent with the formation of a moiré superlattice between the hBN and an underlying highly ordered pyrolytic graphite (HOPG) substrate. This variation is attributed to a periodc modulation of the local density of states and occurs for both exfoliated hBN barriers and epitaxially grown layers. The epitaxial barriers also exhibit enhanced conductance at localized subnanometer regions which are attributed to exposure of the substrate to a nitrogen plasma source during the high temperature growth process. Our results show clearly a spatial periodicity of tunnel current due to the formation of a moiré superlattice and we argue that this can provide a mechanism for elastic scattering of charge carriers for similar interfaces embedded in graphene/hBN resonant tunnel diodes.

Moiré-Modulated Conductance of Hexagonal Boron Nitride Tunnel Barriers.

Hexagonal boron nitride (hBN) has attracted a great deal of attention as a key component in van der Waals (vdW) heterostructures, and as a wide band gap material for deep-ultraviolet devices. We have recently demonstrated plasma-assisted molecular beam epitaxy (PA-MBE) of hBN layers on substrates of highly oriented pyrolytic graphite at high substrate temperatures of ~1400 &deg;C. The current paper will present data on the high-temperature PA-MBE growth of hBN layers using a high-efficiency radio-frequency (RF) nitrogen plasma source. Despite more than a three-fold increase in nitrogen flux with this new source, we saw no significant increase in the growth rates of the hBN layers, indicating that the growth rate of hBN layers is controlled by the boron arrival rate. The hBN thickness increases to 90 nm with decrease in the growth temperature to 1080 &deg;C. However, the decrease in the MBE temperature led to a deterioration in the optical properties of the hBN. The optical absorption data indicates that an increase in the active nitrogen flux during the PA-MBE process improves the optical properties of hBN and suppresses defect related optical absorption in the energy range 5.0⁻5.5 eV.

High-Temperature Molecular Beam Epitaxy of Hexagonal Boron Nitride with High Active Nitrogen Fluxes.

Single-photon emitters (SPEs) in hexagonal boron nitride (hBN) have garnered increasing attention over the last few years due to their superior optical properties. However, despite the vast range of experimental results and theoretical calculations, the defect structure responsible for the observed emission has remained elusive. Here, by controlling the incorporation of impurities into hBN via various bottom-up synthesis methods and directly through ion implantation, we provide direct evidence that the visible SPEs are carbon related. Room-temperature optically detected magnetic resonance is demonstrated on ensembles of these defects. We perform ion-implantation experiments and confirm that only carbon implantation creates SPEs in the visible spectral range. Computational analysis of the simplest 12 carbon-containing defect species suggest the negatively charged [Formula: see text] defect as a viable candidate and predict that out-of-plane deformations make the defect environmentally sensitive. Our results resolve a long-standing debate about the origin of single emitters at the visible range in hBN and will be key to the deterministic engineering of these defects for quantum photonic devices.

Identifying carbon as the source of visible single-photon emission from hexagonal boron nitride.

Transparent conductive oxides are appealing materials for optoelectronic and plasmonic applications as, amongst other advantages, their properties can be modulated by engineering their defects. Optimisation of this adjustment is, however, a complex design problem. This work examined the modification of the carrier transport properties of sputtered tin-doped indium oxide (ITO) via laser annealing in reactive environments. We relate the optical modifications to the structural, compositional, and electronic properties to elucidate the precise mechanisms behind the reactive laser annealing (ReLA) process. For sufficiently high laser fluence, we reveal an ambient-dependent and purely compositional modulation of the carrier concentration of ITO thin films. Hereby, we demonstrate that ReLA utilises the precise energy delivery of photonic processing to enhance the carrier mobility and finely tune the carrier concentration without significantly affecting the crystal structure. Exploitation of this phenomena may enable one to selectively engineer the optoelectronic properties of ITO, promising an alternative to the exploration of new materials for optoelectronic and photonic applications.

Photo-engineered optoelectronic properties of indium tin oxide via reactive laser annealing.

Ferritin is a protein that stores and releases iron to prevent diseases associated with iron dysregulation in plants, animals, and bacteria. The conversion between iron-loaded holo-ferritin and empty apo-ferritin is an important process for iron regulation. To date, studies of ferritin have used either ensemble measurements to quantify the characteristics of a large number of proteins or single-molecule approaches to interrogate labeled or modified proteins. Here we demonstrate the first real-time study of the dynamics of iron ion loading and biomineralization within a single, unlabeled ferritin protein. Using optical nanotweezers, we trapped single apo- and holo-ferritins indefinitely, distinguished one from the other, and monitored their structural dynamics in real time. The study presented here deepens the understanding of the iron uptake mechanism of ferritin proteins, which may lead to new therapeutics for iron-related diseases.

Optical Monitoring of  Iron Loading into Single, Native Ferritin Proteins.

2D semiconductors (2SEM) can transform many sectors, from information and communication technology to healthcare. To date, top-down approaches to their fabrication, such as exfoliation of bulk crystals by "scotch-tape," are widely used, but have limited prospects for precise engineering of functionalities and scalability. Here, a bottom-up technique based on epitaxy is used to demonstrate high-quality, wafer-scale 2SEM based on the wide band gap gallium selenide (GaSe) compound. GaSe layers of well-defined thickness are developed using a bespoke facility for the epitaxial growth and in situ studies of 2SEM. The dominant centrosymmetry and stacking of the individual van der Waals layers are verified by theory and experiment; their optical anisotropy and resonant absorption in the UV spectrum are exploited for photon sensing in the technological UV-C spectral range, offering a scalable route to deep-UV optoelectronics.

Wafer-Scale Two-Dimensional Semiconductors for Deep UV Sensing.

Cathodoluminescence (CL) spectroscopy is a suitable technique for studying the luminescent properties of optoelectronic materials because CL has no limitation on the excitable bandgap energy and eliminates ambiguous signals due to simple light scattering and resonant Raman scattering potentially involved in the photoluminescence spectra. However, direct CL measurements of atomically thin two-dimensional materials have been difficult due to the small excitation volume that interacts with high-energy electron beams. Herein, distinct CL signals from a monolayer hexagonal BN (hBN), namely mBN, epitaxial film grown on a graphite substrate are shown by using a CL system capable of large-area and surface-sensitive excitation. Spatially resolved CL spectra at 13 K exhibited a predominant 5.5-eV emission band, which has been ascribed to originate from multilayered aggregates of hBN, markedly at thicker areas formed on the step edges of the substrate. Conversely, a faint peak at 6.04 ± 0.01 eV was routinely observed from atomically flat areas, which is assigned as being due to the recombination of phonon-assisted direct excitons of mBN. The CL results support the transition from indirect bandgap in bulk hBN to direct bandgap in mBN. The results also encourage one to elucidate emission properties of other low-dimensional materials by using the present CL configuration.

Cathodoluminescence spectroscopy of monolayer hexagonal boron nitride.

Two-dimensional semiconductors (2DSEM) based on van der Waals crystals offer important avenues for nanotechnologies beyond the constraints of Moore's law and traditional semiconductors, such as silicon (Si). However, their application necessitates precise engineering of material properties and scalable manufacturing processes. The ability to oxidize Si to form silicon dioxide (SiO) was crucial for the adoption of Si in modern technologies. Here, we report on the thermal oxidation of the 2DSEM gallium selenide (GaSe). The nanometer-thick layers are grown by molecular beam epitaxy on transparent sapphire (AlO) and feature a centro-symmetric polymorph of GaSe. Thermal annealing of the layers in an oxygen-rich environment promotes the chemical transformation and full conversion of GaSe into a thin layer of crystalline GaO, paralleled by the formation of coherent GaO/AlO interfaces. Versatile functionalities are demonstrated in photon sensors based on GaSe and GaO, ranging from electrical insulation to unfiltered deep ultraviolet optoelectronics, unlocking the technological potential of GaSe nanostructures and their amorphous and crystalline oxides.

Article	Year
Probing the vapor-liquid phase behaviors of near-critical and supercritical fluids using a shear mode piezoelectric sensor. Analytical chemistry\| PubMed ID: 12585473	2003
High broad-band photoresponsivity of mechanically formed InSe-graphene van der Waals heterostructures. Advanced materials (Deerfield Beach, Fla.)\| PubMed ID: 25981798	2015
Strain-Engineered Graphene Grown on Hexagonal Boron Nitride by Molecular Beam Epitaxy. Scientific reports\| PubMed ID: 26928710	2016
Hexagonal Boron Nitride Tunnel Barriers Grown on Graphite by High Temperature Molecular Beam Epitaxy. Scientific reports\| PubMed ID: 27681943	2016
Lattice-Matched Epitaxial Graphene Grown on Boron Nitride. Nano letters\| PubMed ID: 29211487	2018
Moiré-Modulated Conductance of Hexagonal Boron Nitride Tunnel Barriers. Nano letters\| PubMed ID: 29913062	2018
High-Temperature Molecular Beam Epitaxy of Hexagonal Boron Nitride with High Active Nitrogen Fluxes. Materials (Basel, Switzerland)\| PubMed ID: 29966333	2018
Identifying carbon as the source of visible single-photon emission from hexagonal boron nitride. Nature materials\| PubMed ID: 33139892	2021
Photo-engineered optoelectronic properties of indium tin oxide via reactive laser annealing. Scientific reports\| PubMed ID: 36056036	2022
Optical Monitoring of Iron Loading into Single, Native Ferritin Proteins. Nano letters\| PubMed ID: 37053043	2023
Wafer-Scale Two-Dimensional Semiconductors for Deep UV Sensing. Small (Weinheim an der Bergstrasse, Germany)\| PubMed ID: 37798672	2024
Cathodoluminescence spectroscopy of monolayer hexagonal boron nitride. Scientific reports\| PubMed ID: 38167439	2024
Thin GaO Layers by Thermal Oxidation of van der Waals GaSe Nanostructures for Ultraviolet Photon Sensing. ACS applied nano materials\| PubMed ID: 39144400	2024

Christopher J. Mellor

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