Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy

Fluorescence light microscopy allows multicolor visualization of cellular components with high specificity, but its utility has until recently been constrained by the intrinsic limit of spatial resolution. We applied three-dimensional structured illumination microscopy (3D-SIM) to circumvent this limit and to study the mammalian nucleus. By simultaneously imaging chromatin, nuclear lamina, and the nuclear pore complex (NPC), we observed several features that escape detection by conventional microscopy. We could resolve single NPCs that colocalized with channels in the lamin network and peripheral heterochromatin. We could differentially localize distinct NPC components and detect double-layered invaginations of the nuclear envelope in prophase as previously seen only by electron microscopy. Multicolor 3D-SIM opens new and facile possibilities to analyze subcellular structures beyond the diffraction limit of the emitted light.     

Comparing high density LIDAR and medium resolution GPS generated elevation data for predicting yield stability

High density light detection and ranging (LIDAR) imaging has been shown to be able to define yield stability areas of a field for multi-cropping. Since LIDAR imaging is expensive and not widely available, it was hypothesized that medium resolution GPS elevation data which is commonly collected with variable rate technology (VRT) controllers and crop yield monitors could be used in lieu of LIDAR imaging. If proven, growers would be able to construct yield stability maps of their fields without the expense of obtaining LIDAR imaging. After substituting medium resolution GPS elevation data derived from the crop yield monitors, the procedure developed for developing a crop yield stability map was invoked and tested. The hypothesis that medium resolution GPS data could be used in lieu of LIDAR data was found to be invalid as the map generated incorrectly identified both high yield and medium yield areas of the field as low yielding areas as well as the inverse. While disappointing, high resolution GPS data from real-time kinetics (RTK) systems is yet to be tested and may offer an additional avenue to developing crop yield stability maps.     

Video-rate molecular imaging in vivo with stimulated Raman scattering

Optical imaging in vivo with molecular specificity is important in biomedicine because of its high spatial resolution and sensitivity compared with magnetic resonance imaging. Stimulated Raman scattering (SRS) microscopy allows highly sensitive optical imaging based on vibrational spectroscopy without adding toxic or perturbative labels. However, SRS imaging in living animals and humans has not been feasible because light cannot be collected through thick tissues, and motion-blur arises from slow imaging based on backscattered light. In this work, we enable in vivo SRS imaging by substantially enhancing the collection of the backscattered signal and increasing the imaging speed by three orders of magnitude to video rate. This approach allows label-free in vivo imaging of water, lipid, and protein in skin and mapping of penetration pathways of topically applied drugs in mice and humans.     

Subparticle ultrafast spectrum imaging in 4D electron microscopy

Single-particle imaging of structures has become a powerful methodology in nanoscience and molecular and cell biology. We report the development of subparticle imaging with space, time, and energy resolutions of nanometers, femtoseconds, and millielectron volts, respectively. By using scanning electron probes across optically excited nanoparticles and interfaces, we simultaneously constructed energy-time and space-time maps. Spectrum images were then obtained for the nanoscale dielectric fields, with the energy resolution set by the photon rather than the electron, as demonstrated here with two examples (silver nanoparticles and the metallic copper-vacuum interface). This development thus combines the high spatial resolution of electron microscopy with the high energy resolution of optical techniques and ultrafast temporal response, opening the door to various applications in elemental analysis as well as mapping of interfaces and plasmonics.     

Atomic-resolution microscopy in water

The scanning tunneling microscope is revolutionizing the study of surfaces. In ultra-high vacuum it is capable not only of imaging individual atoms but also of determining energy states on an atom-by-atom basis. It is now possible to operate this instrument in water. Aqueous optical microscopy is confined to a lateral resolution limit of about 2000 angstroms, and aqueous x-ray microscopy has yielded a lateral resolution of 75 angstroms. With a scanning tunneling microscope, an image of a graphite surface immersed in deionized water was obtained with features less than 3 angstroms apart clearly resolved. Further, an image measured in saline solution demonstrated that the instrument can be operated under conditions useful for many biological samples.     

根系分区交替滴灌对棉花产量和水分利用效率的影响

 为了探讨旱区农业节水新途径，在甘肃省石羊河流域下游荒漠绿洲区采用常规滴灌和分根区交替滴灌方式研究不同灌溉方式对大田棉花生长发育、产量、水分利用效率以及土壤水分分布的影响。结果表明，交替滴灌处理棉花叶片气孔开度减小，减少了奢侈的蒸腾损失，灌溉定额较小时根系分区交替滴灌对棉花的株高抑制作用较明显，灌水定额较大时限制作用不显著。根系分区交替滴灌技术在大田条件下可使籽棉产量比常规滴灌处理提高21.1%，总水分利用效率和灌溉水利用效率分别提高17.9%和20.9%。同等产量水平下与常规滴灌相比，交替滴灌可节省30.8%的灌水量。本研究表明根系分区交替滴灌是一种切实可行的节水灌溉技术，可在干旱缺水的棉花生产地区进一步研究和推广应用。     

Three-dimensional readout of flash x-ray images of living sperm in water by atomic-force microscopy

The imaging of living specimens in water by x-ray microscopy can be greatly enhanced with the use of an intense flash x-ray source and sophisticated technologies for reading x-ray images. A subnanosecond [corrected] x-ray pulse from a laser-produced plasma was used to record the x-ray image of living sea urchin sperm in an x-ray resist. The resist relief was visualized at high resolution by atomic-force microscopy. Internal structure of the sperm head was evident, and the carbon density in a flagellum was estimated from the relief height.     

Near-field microscopy through a SiC superlens

The wave nature of light limits the spatial resolution in classical microscopy to about half of the illumination wavelength. Recently, a new approach capable of achieving subwavelength spatial resolution, called superlensing, was invented, challenging the already established method of scanning near-field optical microscopy (SNOM). We combine the advantages of both techniques and demonstrate a novel imaging system where the objects no longer need to be in close proximity to a near-field probe, allowing for optical near-field microscopy of subsurface objects at sub-wavelength-scale lateral resolution.     

Atomic resolution imaging of a carbon nanotube from diffraction intensities

Atomic imaging of three-dimensional structures has required a crystal in diffraction or a lens in electron imaging. Whereas diffraction achieves very high resolution by averaging over many cells, imaging gives localized structural information, such as the position of a single dopant atom. However, lens aberrations limit electron imaging resolution to about 1 angstrom. Resolution is reduced further by low contrast from weakscattering or from the limitations on electron dose for radiation-sensitive molecules. We show that both high resolution and high contrast can be achieved by imaging from diffraction with a nanometer-sized coherent electron beam. The phase problem is solved by oversampling and iterative phase retrieval. We apply this technique to image a double-wall carbon nanotube at 1-angstrom resolution, revealing the structure of two tubes of different helicities. Because the only requirement for imaging is a diffraction pattern sampled below the Nyquist frequency, our technique has the potential to image nonperiodic nanostructures, including biological macromolecules, at diffraction intensity-limited resolutions.     

Asymmetric illumination contrast: a method of image formation for video light microscopy

Images with high resolution and exceptionally broad gray scale can be obtained by the application of video contrast enhancement to an optimized procedure for imaging transparent objects with oblique rays of illumination. This technique is simple to set up. A conventional microscope with a light source whose position can be adjusted and a video camera with controls for gain and black level are the only essential components. Features such as high resolution, optical sectioning, control of contrast, and operation under low light intensity make this technique preferable, in several instances, to currently used video microscopy techniques.     

Imaging of transient structures using nanosecond in situ TEM

The microstructure and properties of a material depend on dynamic processes such as defect motion, nucleation and growth, and phase transitions. Transmission electron microscopy (TEM) can spatially resolve these nanoscale phenomena but lacks the time resolution for direct observation. We used a photoemitted electron pulse to probe dynamic events with "snapshot" diffraction and imaging at 15-nanosecond resolution inside of a dynamic TEM. With the use of this capability, the moving reaction front of reactive nanolaminates is observed in situ. Time-resolved images and diffraction show a transient cellular morphology in a dynamically mixing, self-propagating reaction front, revealing brief phase separation during cooling, and thus provide insights into the mechanisms driving the self-propagating high-temperature synthesis.     

Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy

Recent advances in far-field fluorescence microscopy have led to substantial improvements in image resolution, achieving a near-molecular resolution of 20 to 30 nanometers in the two lateral dimensions. Three-dimensional (3D) nanoscale-resolution imaging, however, remains a challenge. We demonstrated 3D stochastic optical reconstruction microscopy (STORM) by using optical astigmatism to determine both axial and lateral positions of individual fluorophores with nanometer accuracy. Iterative, stochastic activation of photoswitchable probes enables high-precision 3D localization of each probe, and thus the construction of a 3D image, without scanning the sample. Using this approach, we achieved an image resolution of 20 to 30 nanometers in the lateral dimensions and 50 to 60 nanometers in the axial dimension. This development allowed us to resolve the 3D morphology of nanoscopic cellular structures.     

High resolution satellite imaging sensors for precision agriculture

The central concept of precision agriculture is to manage within-field soil and crop growth variability for more efficient use of farming inputs. Remote sensing has been an integral part of precision agriculture since the farming technology started developing in the mid to late 1980s. Various types of remote sensors carried on ground-based platforms, manned aircraft, satellites, and more recently, unmanned aircraft have been used for precision agriculture applications. Original satellite sensors, such as Landsat and SPOT, have commonly been used for agricultural applications over large geographic areas since the 1970s, but they have limited use for precision agriculture because of their relatively coarse spatial resolution and long revisit time. Recent developments in high resolution satellite sensors have significantly narrowed the gap in spatial resolution between satellite imagery and airborne imagery. Since the first high resolution satellite sensor IKONOS was launched in 1999, numerous commercial high resolution satellite sensors have become available. These imaging sensors not only provide images with high spatial resolution, but can also repeatedly view the same target area. The high revisit frequency and fast data turnaround time, combined with their relatively large aerial coverage, make high resolution satellite sensors attractive for many applications, including precision agriculture. This article will provide an overview of commercially available high resolution satellite sensors that have been used or have potential for precision agriculture. The applications of these sensors for precision agriculture are reviewed and application examples based on the studies conducted by the author and his collaborators are provided to illustrate how high resolution satellite imagery has been used for crop identification, crop yield variability mapping and pest management. Some challenges and future directions on the use of high resolution satellite sensors and other types of remote sensors for precision agriculture are discussed.     

High-resolution EM of colloidal nanocrystal growth using graphene liquid cells

We introduce a new type of liquid cell for in situ transmission electron microscopy (TEM) based on entrapment of a liquid film between layers of graphene. The graphene liquid cell facilitates atomic-level resolution imaging while sustaining the most realistic liquid conditions achievable under electron-beam radiation. We employ this cell to explore the mechanism of colloidal platinum nanocrystal growth. Direct atomic-resolution imaging allows us to visualize critical steps in the process, including site-selective coalescence, structural reshaping after coalescence, and surface faceting.     

Formation of Chiral Interdigitated Multilayers at the Air-Liquid Interface Through Acid-Base Interactions

Thin interdigitated films composed of a long-chain, water-insoluble chiral acid (p-pentadecylmandelic acid of absolute configuration R) and a water-soluble chiral base (phenylethylamine, R') were constructed at the air-solution interface. The (R, R') structure was characterized to near-atomic resolution by grazing-incidence x-ray diffraction (GIXD). The two diastereomeric systems, (R, R') and (R, S'), demonstrate similar surface pressure-molecular area isotherms, but their structures are completely different on the molecular level, as monitored by GIXD. Complementary data on these two architectures were provided by atomic force microscopy.     

Mapping winter wheat using phenological feature of peak before winter on the North China Plain based on time-series MODIS data

By employing the unique phenological feature of winter wheat extracted from peak before winter(PBW) and the advantages of moderate resolution imaging spectroradiometer(MODIS) data with high temporal resolution and intermediate spatial resolution,a remote sensing-based model for mapping winter wheat on the North China Plain was built through integration with Landsat images and land-use data. First,a phenological window,PBW was drawn from time-series MODIS data. Next,feature extraction was performed for the PBW to reduce feature dimension and enhance its information. Finally,a regression model was built to model the relationship of the phenological feature and the sample data. The amount of information of the PBW was evaluated and compared with that of the main peak(MP). The relative precision of the mapping reached up to 92% in comparison to the Landsat sample data,and ranged between 87 and 96% in comparison to the statistical data. These results were sufficient to satisfy the accuracy requirements for winter wheat mapping at a large scale. Moreover,the proposed method has the ability to obtain the distribution information for winter wheat in an earlier period than previous studies. This study could throw light on the monitoring of winter wheat in China by using unique phenological feature of winter wheat.     

Scanning tunneling microscopy of galena (100) surface oxidation and sorption of aqueous gold

Scanning tunneling microscopy was used to characterize the growth of oxidized areas on galena (100) surfaces and the formation of gold islands by the reductive adsorption of AuCl(4)(-) from aqueous solution. The gold islands and galena substrate were distinguished by atomic resolution imaging and tunneling spectroscopy. Oxidized areas on galena have [110]-trending boundaries; gold islands elongate along [110] directions. However, there are no obvious structural registry considerations that would lead to elongation of gold islands in a [110] direction. Instead, it is probable that a direct coupling of gold reduction and sulfide surface oxidation controls the initial formation of gold islands. Gold islands grow less quickly on preoxidized galena surfaces and show no preferred direction of growth.     

Multicolor super-resolution imaging with photo-switchable fluorescent probes

Recent advances in far-field optical nanoscopy have enabled fluorescence imaging with a spatial resolution of 20 to 50 nanometers. Multicolor super-resolution imaging, however, remains a challenging task. Here, we introduce a family of photo-switchable fluorescent probes and demonstrate multicolor stochastic optical reconstruction microscopy (STORM). Each probe consists of a photo-switchable "reporter" fluorophore that can be cycled between fluorescent and dark states, and an "activator" that facilitates photo-activation of the reporter. Combinatorial pairing of reporters and activators allows the creation of probes with many distinct colors. Iterative, color-specific activation of sparse subsets of these probes allows their localization with nanometer accuracy, enabling the construction of a super-resolution STORM image. Using this approach, we demonstrate multicolor imaging of DNA model samples and mammalian cells with 20- to 30-nanometer resolution. This technique will facilitate direct visualization of molecular interactions at the nanometer scale.     

Scanning interferometric apertureless microscopy: optical imaging at 10 angstrom resolution

Interferometric near-field optical microscopy achieving a resolution of 10 angstroms is demonstrated. The scattered electric field variation caused by a vibrating probe tip in close proximity to a sample surface is measured by encoding it as a modulation in the optical phase of one arm of an interferometer. Unlike in regular near-field optical microscopes, where the contrast results from a weak source (or aperture) dipole interacting with the polarizability of the sample, the present form of imaging relies on a fundamentally different contrast mechanism: sensing the dipole-dipole coupling of two externally driven dipoles (the tip and sample dipoles) as their spacing is modulated.