Probing the electromagnetic field of a 15-nanometre hotspot by single molecule imaging

When light illuminates a rough metallic surface, hotspots can appear, where the light is concentrated on the nanometre scale, producing an intense electromagnetic field. This phenomenon, called the surface enhancement effect, has a broad range of potential applications, such as the detection of weak chemical signals. Hotspots are believed to be associated with localized electromagnetic modes, caused by the randomness of the surface texture. Probing the electromagnetic field of the hotspots would offer much insight towards uncovering the mechanism generating the enhancement; however, it requires a spatial resolution of 1-2?nm, which has been a long-standing challenge in optics. The resolution of an optical microscope is limited to about half the wavelength of the incident light, approximately 200-300?nm. Although current state-of-the-art techniques, including near-field scanning optical microscopy, electron energy-loss spectroscopy, cathode luminescence imaging and two-photon photoemission imaging have subwavelength resolution, they either introduce a non-negligible amount of perturbation, complicating interpretation of the data, or operate only in a vacuum. As a result, after more than 30 years since the discovery of the surface enhancement effect, how the local field is distributed remains unknown. Here we present a technique that uses Brownian motion of single molecules to probe the local field. It enables two-dimensional imaging of the fluorescence enhancement profile of single hotspots on the surfaces of aluminium thin films and silver nanoparticle clusters, with accuracy down to 1.2?nm. Strong fluorescence enhancements, up to 54 and 136 times respectively, are observed in those two systems. This strong enhancement indicates that the local field, which decays exponentially from the peak of a hotspot, dominates the fluorescence enhancement profile.     

Lensless imaging of magnetic nanostructures by X-ray spectro-holography

Our knowledge of the structure of matter is largely based on X-ray diffraction studies of periodic structures and the successful transformation (inversion) of the diffraction patterns into real-space atomic maps. But the determination of non-periodic nanoscale structures by X-rays is much more difficult. Inversion of the measured diffuse X-ray intensity patterns suffers from the intrinsic loss of phase information, and direct imaging methods are limited in resolution by the available X-ray optics. Here we demonstrate a versatile technique for imaging nanostructures, based on the use of resonantly tuned soft X-rays for scattering contrast and the direct Fourier inversion of a holographically formed interference pattern. Our implementation places the sample behind a lithographically manufactured mask with a micrometre-sized sample aperture and a nanometre-sized hole that defines a reference beam. As an example, we have used the resonant X-ray magnetic circular dichroism effect to image the random magnetic domain structure in a Co/Pt multilayer film with a spatial resolution of 50 nm. Our technique, which is a form of Fourier transform holography, is transferable to a wide variety of specimens, appears scalable to diffraction-limited resolution, and is well suited for ultrafast single-shot imaging with coherent X-ray free-electron laser sources.     

Non-destructive determination of local strain with 100-nanometre spatial resolution

Structure sizes of approximately 180 nm are now standard in microelectronics, and state-of-the-art fabrication techniques can reduce these to just a few tens of nanometres. But at these length scales, the strain induced at interfaces can locally distort the crystal lattice, which may in turn affect device performance in an unpredictable way. A means of non-destructively characterizing such strain fields with high spatial resolution and sensitivity is therefore highly desirable. One approach is to use Raman spectroscopy, but this is limited by the intrinsic approximately 0.5-microm resolution limit of visible light probes. Techniques based on electron-beam diffraction can achieve the desired nanometre-scale resolution. But either they require complex sample preparation procedures (which may alter the original strain field) or they are sensitive to distortional (but not dilational) strain within only the top few tens of nanometres of the sample surface. X-rays, on the other hand, have a much greater penetration depth, but have not hitherto achieved strain analysis with sub-micrometre resolution. Here we describe a magnifying diffraction imaging procedure for X-rays which achieves a spatial resolution of 100nm in one dimension and a sensitivity of 10(-4) for relative lattice variations. We demonstrate the suitability of this procedure for strain analysis by measuring the strain depth profiles beneath oxidized lines on silicon crystals.     

Schmidt和Angel结构的龙虾眼X射线成像系统性能对比研究

Schmidt型和Angel型的X射线成像系统通过仿生龙虾眼睛微通道结构实现全反射聚焦成像,相比传统的X射线成像方式,具有大视场、高空间分辨率和能量获取能力。研究两个龙虾眼基础结构模型,根据X射线掠入射反射理论,分析对比了Angel和Schmidt结构光线入射焦面时的分布和空间分辨率,并用tracepro软件建立模型仿真验证了两种结构龙虾眼在聚焦和成像时的接收光/发射光、最大值/十字臂、最大值/本底、十字臂/本底多个性能参数。理论分析和仿真结果表明,当龙虾眼透镜焦距较长和通道长宽比大约为50时,由于多次反射时的聚焦及成像,Schmidt模型比Angel模型的信噪比等主要参数高,空间分辨率低;当龙虾眼透镜焦距较短和通道长宽比大约为10时,由于单次反射时的聚焦及成像,Schmidt模型比Angel模型的信噪比等主要参数低,空间分辨率几乎一样。     

Information extraction and CT reconstruction of liver images based on diffraction enhanced imaging

X-ray phase-contrast imaging (PCI) is a new emerging imaging technique that generates a high spatial resolution and high contrast of biological soft tissues compared to conventional radiography. Herein a biomedical application of diffraction enhanced imaging (DEI) is presented. As one of the PCI methods, DEI derives contrast from many different kinds of sample information, such as the sample's X-ray absorption, refraction gradient and ultra-small-angle X-ray scattering (USAXS) properties, and the sample information is expressed by three parametric images. Combined with computed tomography (CT), DEI-CT can produce 3D volumetric images of the sample and can be used for investigating micro-structures of biomedical samples. Our DEI experiments for liver samples were implemented at the topography station of Beijing Synchrotron Radiation Facility (BSRF). The results show that by using our provided information extraction method and DEI-CT reconstruction approach, the obtained parametric images clearly display the inner structures of liver tissues and the morphology of blood vessels. Furthermore, the reconstructed 3D view of the liver blood vessels exhibits the micro blood vessels whose minimum diameter is on the order of about tens of microns, much better than its conventional CT reconstruction at a millimeter resolution. In conclusion, both the information extraction method and DEI-CT have the potential for use in biomedical micro-structures analysis.     

Clathrate nanostructures for mass spectrometry

The ability of mass spectrometry to generate intact biomolecular ions efficiently in the gas phase has led to its widespread application in metabolomics, proteomics, biological imaging, biomarker discovery and clinical assays (namely neonatal screens). Matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization have been at the forefront of these developments. However, matrix application complicates the use of MALDI for cellular, tissue, biofluid and microarray analysis and can limit the spatial resolution because of the matrix crystal size (typically more than 10 mum), sensitivity and detection of small compounds (less than 500 Da). Secondary-ion mass spectrometry has extremely high lateral resolution (100 nm) and has found biological applications although the energetic desorption/ionization is a limitation owing to molecular fragmentation. Here we introduce nanostructure-initiator mass spectrometry (NIMS), a tool for spatially defined mass analysis. NIMS uses 'initiator' molecules trapped in nanostructured surfaces or 'clathrates' to release and ionize intact molecules adsorbed on the surface. This surface responds to both ion and laser irradiation. The lateral resolution (ion-NIMS about 150 nm), sensitivity, matrix-free and reduced fragmentation of NIMS allows direct characterization of peptide microarrays, direct mass analysis of single cells, tissue imaging, and direct characterization of blood and urine.     

合肥光源X射线成像光束线和实验站设计

在X射线成像光束线光学系统的设计基础上,对光束线的能量分辨率和光子通量进行了计算,并对双晶单色器第一块晶体的热载影响进行了模拟分析,同时利用波带片成像技术对实验站进行了设计,达到了60 nm的空间分辨率.结果表明光束线和实验站的设计能够满足X射线成像实验所需条件的要求.     

Nanoscale imaging magnetometry with diamond spins under ambient conditions

Magnetic resonance imaging and optical microscopy are key technologies in the life sciences. For microbiological studies, especially of the inner workings of single cells, optical microscopy is normally used because it easily achieves resolution close to the optical wavelength. But in conventional microscopy, diffraction limits the resolution to about half the wavelength. Recently, it was shown that this limit can be partly overcome by nonlinear imaging techniques, but there is still a barrier to reaching the molecular scale. In contrast, in magnetic resonance imaging the spatial resolution is not determined by diffraction; rather, it is limited by magnetic field sensitivity, and so can in principle go well below the optical wavelength. The sensitivity of magnetic resonance imaging has recently been improved enough to image single cells, and magnetic resonance force microscopy has succeeded in detecting single electrons and small nuclear spin ensembles. However, this technique currently requires cryogenic temperatures, which limit most potential biological applications. Alternatively, single-electron spin states can be detected optically, even at room temperature in some systems. Here we show how magneto-optical spin detection can be used to determine the location of a spin associated with a single nitrogen-vacancy centre in diamond with nanometre resolution under ambient conditions. By placing these nitrogen-vacancy spins in functionalized diamond nanocrystals, biologically specific magnetofluorescent spin markers can be produced. Significantly, we show that this nanometre-scale resolution can be achieved without any probes located closer than typical cell dimensions. Furthermore, we demonstrate the use of a single diamond spin as a scanning probe magnetometer to map nanoscale magnetic field variations. The potential impact of single-spin imaging at room temperature is far-reaching. It could lead to the capability to probe biologically relevant spins in living cells.     

Achromatic Fresnel optics for wideband extreme-ultraviolet and X-ray imaging

Advances in extreme-ultraviolet (EUV) and X-ray optics are providing powerful new capabilities in high-resolution imaging and trace-element analysis of microscopic specimens, and the potential for fabricating devices of smaller critical dimensions in next-generation integrated circuit lithography. However, achieving the highest resolution with such optics usually requires the illuminating EUV or X-ray beam to be highly monochromatic. It would therefore be highly desirable to have large-field-of-view, sub-100-nm resolution optics that are achromatic to a significant degree, allowing more light to be utilized from broader bandwidth sources such as laser-produced plasmas. Here we report an achromatic Fresnel optical system for EUV or X-ray radiation that combines a Fresnel zone plate with a refractive lens with opposite chromatic aberration. We use the large anomalous dispersion property of the refractive lens material near an absorption edge to make its fabrication practical. The resulting structure can deliver a resolution comparable to that of the Fresnel zone plates that have achieved the highest resolution (25 nm; ref. 3) in the entire electromagnetic spectrum, but with an improvement of two or more orders of magnitude in spectral bandwidth.     

High-throughput experimental tools for the materials genome initiative

The materials innovation infrastructure in the materials genome initiative(MGI)consists of three major components:computational tools,experimental tools,and digital data.This article will review experimental tools for high-throughput,high spatial resolution measurements of several materials properties such as elastic modulus,thermal conductivity,specific heat capacity,and thermal expansion.Application of these tools on compositionvarying samples such as diffusion multiples can be used to quickly and efficiently obtain composition–phase–structure–property relationships for materials property database establishment.They can also be used in conjunction with theoretical modeling to find and explain unusual effects to improve the predictability of models.More micron scale resolution experimental tools are in development.These high-throughput tools will be an essential part of MGI.     

Single spin detection by magnetic resonance force microscopy

Magnetic resonance imaging (MRI) is well known as a powerful technique for visualizing subsurface structures with three-dimensional spatial resolution. Pushing the resolution below 1 micro m remains a major challenge, however, owing to the sensitivity limitations of conventional inductive detection techniques. Currently, the smallest volume elements in an image must contain at least 10(12) nuclear spins for MRI-based microscopy, or 10(7) electron spins for electron spin resonance microscopy. Magnetic resonance force microscopy (MRFM) was proposed as a means to improve detection sensitivity to the single-spin level, and thus enable three-dimensional imaging of macromolecules (for example, proteins) with atomic resolution. MRFM has also been proposed as a qubit readout device for spin-based quantum computers. Here we report the detection of an individual electron spin by MRFM. A spatial resolution of 25 nm in one dimension was obtained for an unpaired spin in silicon dioxide. The measured signal is consistent with a model in which the spin is aligned parallel or anti-parallel to the effective field, with a rotating-frame relaxation time of 760 ms. The long relaxation time suggests that the state of an individual spin can be monitored for extended periods of time, even while subjected to a complex set of manipulations that are part of the MRFM measurement protocol.     

Nanoscale magnetic sensing with an individual electronic spin in diamond

Detection of weak magnetic fields with nanoscale spatial resolution is an outstanding problem in the biological and physical sciences. For example, at a distance of 10 nm, the spin of a single electron produces a magnetic field of about 1 muT, and the corresponding field from a single proton is a few nanoteslas. A sensor able to detect such magnetic fields with nanometre spatial resolution would enable powerful applications, ranging from the detection of magnetic resonance signals from individual electron or nuclear spins in complex biological molecules to readout of classical or quantum bits of information encoded in an electron or nuclear spin memory. Here we experimentally demonstrate an approach to such nanoscale magnetic sensing, using coherent manipulation of an individual electronic spin qubit associated with a nitrogen-vacancy impurity in diamond at room temperature. Using an ultra-pure diamond sample, we achieve detection of 3 nT magnetic fields at kilohertz frequencies after 100 s of averaging. In addition, we demonstrate a sensitivity of 0.5 muT Hz(-1/2) for a diamond nanocrystal with a diameter of 30 nm.     

Application of synchrotron source based DEI method in guinea pig cochleae imaging

Hard X-ray diffraction enhanced imaging (DEI), which is based on a synchrotron source and monochromator-analyzer-crystal system, is an effective method for imaging X-ray phase shift. Utilizing an analyzer crystal with high angular sensitivity of micro-radian, DEI can measure the transmitted, refracted and scattered X-rays when projecting onto a sample. It dramatically improves the contrast and spatial resolution of the resultant images. At the topography station of Beijing Synchrotron Radiation Facilities (BSRF), we implemented DEI method in guinea pig cochleae imaging and acquired a series of DEI images. Based on these images, the apparent absorption and refraction images were calculated. The DEI images revealed the holistic spiral structures and inner details of guinea pig cochleae clearly, even including the structures at the cellular level, such as the static cilia of hairy cells and the limbus of Hansen cell. Due to the advantages of high contrast, high spatial resolution and distinct edge-enhanced effect, DEI method promises extensive applications in biology, medicine and clinic in the near future.     

Application of synchrotron source based DEI method in guinea pig cochleae imaging

Hard X-ray diffraction enhanced imaging (DEI), which is based on a synchrotron source and monochromator-analyzer-crystal system, is an effective method for imaging X-ray phase shift. Utilizing an analyzer crystal with high angular sensitivity of micro-radian, DEI can measure the transmitted, refracted and scattered X-rays when projecting onto a sample. It dramatically improves the contrast and spatial resolution of the resultant images. At the topography station of Beijing Synchrotron Radiation Facilities (BSRF), we implemented DEI method in guinea pig cochleae imaging and acquired a series of DEI images. Based on these images, the apparent absorption and refraction images were calculated. The DEI images revealed the holistic spiral structures and inner details of guinea pig cochleae clearly, even including the structures at the cellular level, such as the static cilia of hairy cells and the limbus of Hansen cell. Due to the advantages of high contrast, high spatial resolution and distinct edge-enhanced effect, DEI method promises extensive applications in biology, medicine and clinic in the near future.     

Probing carrier dynamics in nanostructures by picosecond cathodoluminescence

Picosecond and femtosecond spectroscopy allow the detailed study of carrier dynamics in nanostructured materials. In such experiments, a laser pulse normally excites several nanostructures at once. However, spectroscopic information may also be acquired using pulses from an electron beam in a modern electron microscope, exploiting a phenomenon called cathodoluminescence. This approach offers several advantages. The multimode imaging capabilities of the electron microscope enable the correlation of optical properties (via cathodoluminescence) with surface morphology (secondary electron mode) at the nanometre scale. The broad energy range of the electrons can excite wide-bandgap materials, such as diamond- or gallium-nitride-based structures that are not easily excited by conventional optical means. But perhaps most intriguingly, the small beam can probe a single selected nanostructure. Here we apply an original time-resolved cathodoluminescence set-up to describe carrier dynamics within single gallium-arsenide-based pyramidal nanostructures with a time resolution of 10 picoseconds and a spatial resolution of 50 nanometres. The behaviour of such charge carriers could be useful for evaluating elementary components in quantum computers, optical quantum gates or single photon sources for quantum cryptography.     

Structural studies of membranes and surface layers up to 1,000 A thick using X-ray standing waves

The X-ray standing wave (XSW) method, developed in the 1960s, was used originally to determine heavy atom positions in and on silicon and germanium single crystals. An X-ray standing wave generated by the interference of coherent incident and reflected beams excites X-ray fluorescence from the heavy atom, the intensity of which as a function of incident angle provides an indication of the atom's distance from the X-ray reflecting surface. The availability of X-ray mirrors and the ability to prepare layered synthetic microstructures has made possible the study of biologically relevant structures using the XSW technique on length scales of typically tens to hundreds of ?ngstr?ms, allowing heavy atoms in such structures to be located with ?ngstr?m or sub?ngstr?m resolution. Many model biological systems (such as Langmuir-Blodgett films, which mimic membranes) require access to still larger scales, but it is not obvious that an XSW will remain coherent over such length scales. Here we report studies of a lipid multilayer system using the XSW method, in which we have been able to locate the metal atoms in a zinc arachidate bilayer with ?ngstr?m resolution at a distance of almost 1,000 A above the surface of a gold mirror. Our results indicate that the XSW technique should be useful for structural studies of supramolecular aggregates, receptor-ligand interactions and multi-membrane stacks, in which length scales of this order are encountered.     

Progress of AFM single-cell and single-molecule morphology imaging

Atomic force microscopy (AFM) can probe single living cells and single native membrane proteins in natural fluid environments with label-free high spatial resolution. It has thus become an important tool for cellular and molecular biology that significantly complements traditional biochemical and biophysical techniques such as optical and electron microscopy and X-ray crystallog-raphy. Imaging surface topography is the primary application of AFM in the life sciences. Since the early 1990s, researchers have used AFM to investigate morphological features of living cells and native membrane proteins with impressive results. Steady improvements in AFM techniques for imaging soft biological samples have greatly expanded its applications. Based on the authors’ own research in AFM imaging of living cell morphologies, a review of sample preparation procedures for single-cell and single-molecule imaging experiments is presented, along with a summary of recent progress in AFM imaging of living cells and native membrane proteins. Finally, the challenges of AFM high-resolution imaging at the single-cell and single-molecule levels are discussed.     

Super-resolution microscopy of live cells using single molecule localization

The resolution of conventional light microscopy is insufficient for subcelluar studies. The invention of various super-resolution imaging techniques breaks the diffraction barrier and pushes the resolution limit towards the nanometer scale. Here, we focus on a category of super-resolution microscopy that relies on the stochastic activation and precise localization of single molecules. A diversity of fluorescent probes with different characteristics has been developed to achieve super-resolution imaging. In addition, with the implementation of robust localization algorithms, this family of approaches has been expanded to multi-color, three-dimensional and live cell imaging, which provides a promising prospect in biological research.     

High resolution imaging using scanning ion conductance microscopy with improved distance feedback control

Microscopy is an essential technique for observation on living cells. There is currently great interest in apply scanning probe microscopy to image living biological cells in their natural environment at the nanometer scale. Scanning ion conductance microscopy is a new form of scanning probe microscopy, which enables non-contact high resolution imaging of living biological cells. Based on a scanned nanopipette in physiological buffer, the distance feedback control uses the ion current to control the distance between the pipette tip and the sample surface. However, this feedback control has difficulties over slopes on convoluted cell surfaces, which limits its resolution. In this study, we present an improved form of feedback control that removes the contribution of up to the third order slope from the ion current signal, hence providing a more accurate signal for controlling the distance. We show that this allows faster and lower noise topographic high resolution imaging.     

Three-dimensional X-ray structural microscopy with submicrometre resolution

Advanced materials and processing techniques are based largely on the generation and control of non-homogeneous microstructures, such as precipitates and grain boundaries. X-ray tomography can provide three-dimensional density and chemical distributions of such structures with submicrometre resolution; structural methods exist that give submicrometre resolution in two dimensions; and techniques are available for obtaining grain-centroid positions and grain-average strains in three dimensions. But non-destructive point-to-point three-dimensional structural probes have not hitherto been available for investigations at the critical mesoscopic length scales (tenths to hundreds of micrometres). As a result, investigations of three-dimensional mesoscale phenomena--such as grain growth, deformation, crumpling and strain-gradient effects--rely increasingly on computation and modelling without direct experimental input. Here we describe a three-dimensional X-ray microscopy technique that uses polychromatic synchrotron X-ray microbeams to probe local crystal structure, orientation and strain tensors with submicrometre spatial resolution. We demonstrate the utility of this approach with micrometre-resolution three-dimensional measurements of grain orientations and sizes in polycrystalline aluminium, and with micrometre depth-resolved measurements of elastic strain tensors in cylindrically bent silicon. This technique is applicable to single-crystal, polycrystalline, composite and functionally graded materials.