Protein Nanopore Membranes Prepared by a Simple Langmuir-Schaefer Approach.

Filtration through membranes with nanopores is typically associated with high transmembrane pressures and high energy consumption. This problem can be addressed by reducing the respective membrane thickness. Here, a simple procedure is described to prepare ultrathin membranes based on protein nanopores, which exhibit excellent water permeance, two orders of magnitude superior to comparable, industrially applied membranes. Furthermore, incorporation of either closed or open protein nanopores allows tailoring the membrane's ion permeability. To form such membranes, the transmembrane protein ferric hydroxamate uptake protein component A (FhuA) or its open-pore variant are assembled at the air-water interface of a Langmuir trough, compressed to a dense film, crosslinked by glutaraldehyde, and transferred to various support materials. This approach allows to prepare monolayer or multilayer membranes with a very high density of protein nanopores. Freestanding membranes covering holes up to 5 µm in diameter are visualized by atomic force microscopy (AFM), helium ion microscopy, and transmission electron microscopy. AFM PeakForce quantitative nanomechanical property mapping (PeakForce QNM)  demonstrates remarkable mechanical stability and elastic properties of freestanding monolayer membranes with a thickness of only 5 nm. The new protein membrane can pave the way to energy-efficient nanofiltration.

Surface-Enhanced Raman Spectroscopy of Carbon Nanomembranes from Aromatic Self-Assembled Monolayers.

Surface-enhanced Raman scattering spectroscopy (SERS) was employed to investigate the formation of self-assembled monolayers (SAMs) of biphenylthiol, 4'-nitro-1,1'-biphenyl-4-thiol, and p-terphenylthiol on Au surfaces and their structural transformations into carbon nanomembranes (CNMs) induced by electron irradiation. The high sensitivity of SERS allows us to identify two types of Raman scattering in electron-irradiated SAMs: (1) Raman-active sites exhibit similar bands as those of pristine SAMs in the fingerprint spectral region, but with indications of an amorphization process and (2) Raman-inactive sites show almost no Raman-scattering signals, except a very weak and broad D band, indicating a lack of structural order but for the presence of graphitic domains. Statistical analysis showed that the ratio of the number of Raman-active sites to the total number of measurement sites decreases exponentially with increasing the electron irradiation dose. The maximum degree of cross-linking ranged from 97 to 99% for the three SAMs. Proof-of-concept experiments were conducted to demonstrate potential applications of Raman-inactive CNMs as a supporting membrane for Raman analysis.

Helium Ion Microscopy Visualizes Lipid Nanodomains in Mammalian Cells.

Cell membranes are composed of 2D bilayers of amphipathic lipids, which allow a lateral movement of the respective membrane components. These components are arranged in an inhomogeneous manner as transient micro- and nanodomains, which are believed to be crucially involved in the regulation of signal transduction pathways in mammalian cells. Because of their small size (diameter 10-200 nm), membrane nanodomains cannot be directly imaged using conventional light microscopy. Here, direct visualization of cell membrane nanodomains by helium ion microscopy (HIM) is presented. It is shown that HIM is capable to image biological specimens without any conductive coating and that HIM images clearly allow the identification of nanodomains in the ultrastructure of membranes with 1.5 nm resolution. The shape of these nanodomains is preserved by fixation of the surrounding unsaturated fatty acids while saturated fatty acids inside the nanodomains are selectively removed. Atomic force microscopy, fluorescence microscopy, 3D structured illumination microscopy, and direct stochastic optical reconstruction microscopy provide additional evidence that the structures in the HIM images of cell membranes originate from membrane nanodomains. The nanodomains observed by HIM have an average diameter of 20 nm and are densely arranged with a minimal nearest neighbor distance of ≈ 15 nm.

Controlled translocation of DNA through nanopores in carbon nano-, silicon-nitride- and lipid-coated membranes.

We investigated experimentally and theoretically the translocation forces when a charged polymer is threaded through a solid-state nanopore and found distinct dependencies on the nanopore diameter as well as on the nano membrane material chemistry. For this purpose we utilized dedicated optical tweezers force mechanics capable of probing the insertion of negatively charged double-stranded DNA inside a helium-ion drilled nanopore. We found that both the diameter of the nanopore and the membrane material itself have significant influences on the electroosmotic flow through the nanopore and thus on the threading force. Compared to a bare silicon-nitride membrane, the threading of DNA through only 3 nm thin carbon nano membranes as well as lipid bilayer-coated nanopores increased the threading force by 15% or 85%, respectively. This finding was quantitatively described by our recently developed theoretical model that also incorporates hydrodynamic slip effects on the translocating DNA molecule and the force dependence on the membrane thickness. The additional measurements presented in this paper further support our model.

Preparation of conductive gold nanowires in confined environment of gold-filled polymer nanotubes.

Continuous conductive gold nanofibers are prepared via the "tubes by fiber templates" process. First, poly(l-lactide) (PLLA)-stabilized gold nanoparticles (AuNP) with over 60 wt% gold are synthesized and characterized, including gel permeation chromatography coupled with a diode array detector. Subsequent electrospinning of these AuNP with template PLLA results in composite nanofibers featuring a high gold content of 57 wt%. Highly homogeneous gold nanowires are obtained after chemical vapor deposition of 345 nm of poly(p-xylylene) (PPX) onto the composite fibers followed by pyrolysis of the polymers at 1050 °C. The corresponding heat-induced transition from continuous gold-loaded polymer tubes to smooth gold nanofibers is studied by transmission electron microscopy and helium ion microscopy using both secondary electrons and Rutherford backscattered ions.

Toward plasmonics with nanometer precision: nonlinear optics of helium-ion milled gold nanoantennas.

Plasmonic nanoantennas are versatile tools for coherently controlling and directing light on the nanoscale. For these antennas, current fabrication techniques such as electron beam lithography (EBL) or focused ion beam (FIB) milling with Ga(+)-ions routinely achieve feature sizes in the 10 nm range. However, they suffer increasingly from inherent limitations when a precision of single nanometers down to atomic length scales is required, where exciting quantum mechanical effects are expected to affect the nanoantenna optics. Here, we demonstrate that a combined approach of Ga(+)-FIB and milling-based He(+)-ion lithography (HIL) for the fabrication of nanoantennas offers to readily overcome some of these limitations. Gold bowtie antennas with 6 nm gap size were fabricated with single-nanometer accuracy and high reproducibility. Using third harmonic (TH) spectroscopy, we find a substantial enhancement of the nonlinear emission intensity of single HIL-antennas compared to those produced by state-of-the-art gallium-based milling. Moreover, HIL-antennas show a vastly improved polarization contrast. This superior nonlinear performance of HIL-derived plasmonic structures is an excellent testimonial to the application of He(+)-ion beam milling for ultrahigh precision nanofabrication, which in turn can be viewed as a stepping stone to mastering quantum optical investigations in the near-field.

Tailoring the mechanics of ultrathin carbon nanomembranes by molecular design.

Freestanding carbon nanomembranes (CNMs) with a thickness between 0.6 and 1.7 nm were prepared from self-assembled monolayers (SAMs) of diverse polyaromatic precursors via low-energy electron-induced cross-linking. The mechanical properties of CNMs were investigated using AFM bulge test, where a pressure difference was applied to the membrane and the resulting deflection was measured by atomic force microscopy. We found a correlation between the rigidity of the precursor molecules and the macroscopic mechanical stiffness of CNMs. While CNMs from rigid and condensed precursors like naphthalene and pyrene thiols prove to exhibit higher Young's moduli of 15-19 GPa, CNMs from nonfused oligophenyls possess lower Young's moduli of ~10 GPa. For CNMs from less densely packed SAMs, the presence of defects and nanopores plays an important role in determining their mechanical properties. The finite element method (FEM) was applied to examine the deformation profiles and simulate the pressure-deflection relationships.

Enhanced infrared LSPR sensitivity of cap-shaped gold nanoparticles coupled to a metallic film.

We report on optical properties of gold deposited on SiO2 nanospheres randomly adsorbed on a thin gold layer. Extinction peaks with optical density of more than 2 are observed in the visible as well as near-IR regimes. The peak wavelength of the latter was affected exquisitely by the thickness of the top layer. A helium ion microscope (HIM) was used for careful observation of morphological transformation accompanying the change in the deposition thickness. Growth of grain structures into a capped-dimer structure was accompanied by slight blue-shift of the visible peak and significantly greater red-shift of the near-IR peak. Our finite-difference time-domain (FDTD) calculations show that these peaks in the visible and near-IR can be respectively attributed to dipole modes associated with transverse and longitudinal oscillations of free electrons in the gold-capped dimer. To investigate the refractive index sensitivity of these peaks, we used two approaches: immersion in solutions of varying refractive index and coating with an organic layer. With the first approach that characterizes the bulk sensitivity, the visible peak shows sensitivity of 122 nm/RIU, while the near-IR peak shifts at the rate of 506 nm/RIU. With the second approach that reflects the local sensitivity, the surface was saturated with alkaline phosphatase (ALP), whose subsequent reaction led to formation of a thin insoluble organic layer, causing a relatively small blue-shift, under 7 nm, of the visible peak and much larger red-shift, over 50 nm, of the near-IR peak when measured in buffer. When the same reaction was measured at end points in the air, the shift was as large as 444 nm for the near-IR peak.

Defect-Healed Carbon Nanomembranes for Enhanced Salt Separation: Scalable Synthesis and Performance.

Carbon nanomembranes (CNMs), with a high density of subnanometer channels, enable superior salt separation performance compared to conventional membranes. However, defects that occur during the synthesis and transfer processes impede their technical realization on a macroscopic scale. Here, we introduce a practical and scalable interfacial polymerization method to effectively heal defects while preserving the subnanometer pores within CNMs. The defect-healed freestanding CNMs show an exceptional performance in forward osmosis (FO), achieving a water flux of 105 L m-2 h-1 and a specific reverse salt flux of 0.1 g L-1 when measured with 1 M NaCl as draw solution. This water flux is 10 times higher than that of commercially available FO membranes, and the reverse salt flux is 70% lower. Through successful implementation of the defect-healing method and support optimization, we demonstrate the synthesis of fully functional, centimeter-scale CNM-based composite membranes showing high water permeance and a high salt rejection. Our defect-healing method presents a promising pathway to overcome limitations in CNM synthesis, advancing their potential for practical salt separation applications.

Imaging nanocarbon materials: soot particles in flames are not structurally homogeneous.

For the first time, nascent soot particles are probed by using helium-ion microscopy (HIM). HIM is a technique that is similar to scanning electron microscopy (SEM) but it can achieve higher contrast and improved surface sensitivity, especially for carbonaceous materials. The HIM microscope yields images with a high contrast, which allows for the unambiguous recognition of smaller nascent soot particles than those observed in previous transmission electron microscopy studies. The results indicate that HIM is ideal for rapid and reliable probing of the morphology of nascent soot, with surface details visible down to approximately 5 nm, and particles as small as 2 nm are detectable. The results also show that nascent soot is structurally and chemically inhomogeneous, and even the smallest particles can have shapes that deviate from a perfect sphere.

Quantitative high-resolution transmission electron microscopy of single atoms.

Single atoms can be considered as the most basic objects for electron microscopy to test the microscope performance and basic concepts for modeling image contrast. In this work high-resolution transmission electron microscopy was applied to image single platinum, molybdenum, and titanium atoms in an aberration-corrected transmission electron microscope. The atoms are deposited on a self-assembled monolayer substrate that induces only negligible contrast. Single-atom contrast simulations were performed on the basis of Weickenmeier-Kohl and Doyle-Turner form factors. Experimental and simulated image intensities are in quantitative agreement on an absolute intensity scale, which is provided by the vacuum image intensity. This demonstrates that direct testing of basic properties such as form factors becomes feasible.

Mechanically stacked 1-nm-thick carbon nanosheets: ultrathin layered materials with tunable optical, chemical, and electrical properties.

Carbon nanosheets are mechanically stable, free-standing two-dimensional materials with a thickness of ≈1 nm and well defined physical and chemical properties. They are made by radiation-induced cross-linking of aromatic self-assembled monolayers. Herein, a route is presented to the scalable fabrication of multilayer nanosheets with tunable electrical, optical, and chemical properties on insulating substrates. Stacks of up to five nanosheets with sizes of ≈1 cm(2) on oxidized silicon are studied. Their optical characteristics are investigated by visual inspection, optical microscopy, UV-vis reflection spectroscopy, and model calculations. Their chemical composition is studied by X-ray photoelectron spectroscopy. The multilayer samples are then annealed in an ultrahigh vacuum at various temperatures up to 1100 K. A subsequent investigation by Raman, X-ray photoelectron, and UV-vis reflection spectroscopy, as well as by electrical four-point probe measurements, demonstrates that the layered nanosheets transform into nanocrystalline graphene. This structural and chemical transformation is accompanied by changes in the optical properties and electrical conductivity and opens up a new path for the fabrication of ultrathin functional conductive coatings.

Patterned polymer carpets.

For the development of polymer carpets as active devices for micro- and nanotechnology, a control of the polymer carpet morphology and especially control of the stimulus responsive polymer brush is needed. Here, we report on the first example for the fabrication of patterned polymer carpets. On a two-dimensional framework of fully crosslinked and chemically patterned nanosheets, polymer brushes of styrene and 4-vinyl pyridine were grafted by self-initiated surface photopolymerization and photografting (SIPGP). It was found that polymer grafting by SIPGP occurred over the entire nanosheets but with a preferred grafting on the amino functionalized nanosheet areas. This results in continuous polymer carpets with an intact nanosheet framework but with amplification of the chemical patterning into a three dimensional topography of the grafted polymer brush. In the case of negative patterned nanosheets, the patterned carpet could be prepared as freestanding ultrathin membranes. Furthermore, swelling experiments with poly(4-vinyl pyridine) carpets showed that the patterns induces a directional buckling of the flexible polymer carpet. This may open the possibility of the development of micro- or nanoactuator devices with anisotropic responds upon environmental changes.

Mechanics of free-standing inorganic and molecular 2D materials.

The discovery of graphene has triggered a great interest in inorganic as well as molecular two-dimensional (2D) materials. In this review, we summarize recent progress in the mechanical characterization of free-standing 2D materials, such as graphene, hexagonal boron nitride (hBN), transition metal-dichalcogenides, MXenes, black phosphor, carbon nanomembranes (CNMs), 2D polymers, 2D metal organic frameworks (MOFs) and covalent organic frameworks (COFs). Elastic, fracture, bending and interfacial properties of these materials have been determined using a variety of experimental techniques including atomic force microscopy based nanoindentation, in situ tensile/fracture testing, bulge testing, Raman spectroscopy, Brillouin light scattering and buckling-based metrology. Additionally, we address recent advances of 2D materials in a variety of mechanical applications, including resonators, microphones and nanoelectromechanical sensors. With the emphasis on progress and challenges in the mechanical characterization of inorganic and molecular 2D materials, we expect a continuous growth of interest and more systematic experimental work on the mechanics of such ultrathin nanomaterials.

Scanning transmission helium ion microscopy on carbon nanomembranes.

A dark-field scanning transmission ion microscopy detector was designed for the helium ion microscope. The detection principle is based on a secondary electron conversion holder with an exchangeable aperture strip allowing its acceptance angle to be tuned from 3 to 98 mrad. The contrast mechanism and performance were investigated using freestanding nanometer-thin carbon membranes. The results demonstrate that the detector can be optimized either for most efficient signal collection or for maximum image contrast. The designed setup allows for the imaging of thin low-density materials that otherwise provide little signal or contrast and for a clear end-point detection in the fabrication of nanopores. In addition, the detector is able to determine the thickness of membranes with sub-nanometer precision by quantitatively evaluating the image signal and comparing the results with Monte Carlo simulations. The thickness determined by the dark-field transmission detector is compared to X-ray photoelectron spectroscopy and energy-filtered transmission electron microscopy measurements.

Combined nanofiltration and advanced oxidation processes with bifunctional carbon nanomembranes.

Wastewater reclamation is becoming a top global interest as population growth and rapid industrialization pose a major challenge that requires development of sustainable cost-effective technologies and strategies for wastewater treatment. Carbon nanomembranes (CNMs)-synthetic 2D carbon sheets-can be tailored chemically with specific surface functions and/or physically with nanopores of well-defined size as a strategy for multifunctional membrane design. Here, we explore a bifunctional design for combined secondary wastewater effluent treatment with dual action of membrane separation and advanced oxidation processes (AOP), exploiting dissolved oxygen. The bifunctional membrane consists of a CNM layer on top of a commercial ultrafiltration membrane (Microlon™) and a spray-coated reduced graphene oxide (rGO) thin film as the bottom layer. The CNM/support/rGO membrane was characterized by helium ion and atomic force microscopy, FTIR, XPS with a four-point conductivity probe, cyclic voltammetry, galvanostatic measurements, and impedance spectroscopy. Combined treatment of water by nanofiltration and AOP was demonstrated, employing a unique three electrode-dead end filtration setup that enables concurrent application of potential and pressure on the integrated membrane. For the model organic compound methylene blue, oxidation (by the Fenton reaction) was evaluated using UV-vis (610 nm). The rejection rate and permeability provided by the CNM layer were evaluated by dissolving polyethylene glycol (400 and 1000 Da) in the feed solution and applying pressure up to 1.5 bar. This demonstration of combined membrane separation and AOP using an integrated membrane opens up a new strategy for wastewater treatment.

Polymer carpets.

The fabrication of defined polymer objects of reduced dimensions such as polymer-coated nanoparticles (zero-dimensional (0D)), cylindrical brushes (1D), and polymer membranes (2D), is currently the subject of intense research. In particular, ultrathin polymer membranes with high aspect ratios are being discussed as novel materials for miniaturized sensors because they would provide extraordinary sensitivity and dynamic range when sufficient mechanical stability can be combined with flexibility and chemical functionality. Unlike current approaches that rely on crosslinking of polymer layers for stabilization, this report presents the preparation of a new class of polymer material, so-called "polymer carpets," a freestanding polymer brush grown by surface-initiated polymerization on a crosslinked 1-nm-thick monolayer. The solid-supported, as well as freestanding, polymer carpets are found to be mechanically robust and to react instantaneously and reversibly to external stimuli by buckling. The carpet mechanics and the dramatic changes of the film properties (optical, wetting) upon chemical stimuli are investigated in detail as they allow the development of completely new integrated micro-/nanotechnology devices.

Holey nanosheets by patterning with UV/ozone.

Self-assembled monolayers (SAMs) of 1,1'-biphenyl-4-thiol are patterned by using UV/ozone treatment through a shadow mask. After subsequent irradiation with low-energy electrons, the SAM is cross-linked to form a two-dimensional, holey nanosheet with a thickness of approximately 1 nm. This nanosheet is mechanically stable and can be released from the original gold substrate and transferred to new substrates, including transmission electron microscopy (TEM) grids. The process is monitored by complementary microscopic techniques as well as X-ray photoelectron spectroscopy (XPS).

Ultrathin conductive carbon nanomembranes as support films for structural analysis of biological specimens.

Ultrathin carbon nanomembranes (CNM) have been tested as supports for both cryogenic high-resolution transmission electron microscopy (cryo-EM) as well as atomic force microscopy (AFM) of biological specimens. Purple membrane (PM) from Halobacterium salinarum, a 2-D crystalline monolayer of bacteriorhodopsin (BR) and lipids, was used for this study. Due to their low thickness of just 1.6 nm CNM add virtually no phase contrast to the transmission pattern. This is an important advantage over commonly used amorphous carbon support films which become instable below a thickness of approximately 20 nm. Moreover, the electrical conductivity of CNM can be tuned leading to conductive carbon nanomembranes (cCNM). cCNM support films were analyzed for the first time and were found to ideally meet all requirements of cryo-EM of insulating biological samples. A projection map of PM on cCNM at 4 A resolution has been calculated which proves that the structural integrity of biological samples is preserved up to the high-resolution range. CNM have also proven to be suitable supports for AFM analysis of biological samples. PM on CNM was imaged at molecular resolution and single molecule force spectra were recorded which show no differences compared to force spectra of PM obtained with other substrates. This is the first demonstration of a support film material which meets the requirements of both, cryo-EM and AFM, thus enabling comparative structural studies of biomolecular samples with unchanged sample-substrate interactions. Beyond high-resolution cryo-EM of biological samples, cCNM are attractive new substrates for other biophysical techniques which require conductive supports, i.e. scanning tunneling microscopy (STM) and electrostatic force microscopy (EFM).

Selective Diffusion of CO2 and H2O through Carbon Nanomembranes in Aqueous Solution as Studied with Radioactive Tracers.

Nanometer-thin carbon nanomembranes (CNMs) are promising candidates for efficient separation processes due to their thinness and intrinsic well-defined pore structure. This work used radioactive tracer molecules to characterize diffusion of [3H]H2O, [14C]NaHCO3, and [32P]H3PO4 through a p-[1,1',4',1″]-terphenyl-4-thiol (TPT) CNM in aqueous solution. The experimental setup consisted of two microcompartments separated by a CNM-covered micropore. Tracers were added to one compartment and their time-dependent increase in the other compartment was monitored. Occurring concentration polarization and outgassing effects were fully considered using a newly developed mathematical model. Our findings are consistent with previous gas/vapor permeation measurements. The high sensitivity toward a small molecule flow rate enables quantification of diffusion through micron-sized CNMs in aqueous solution. Furthermore, the results allow unambiguous distinction between intact and defective membranes. Even for extremely small membrane areas, this method allows detailed insight into the transmembrane transport properties, which is crucial for the design of 2D-separation membranes.