Ultra-high Spatial Resolution Research Articles

In vivo real-time visualization of tissue blood flow and angiogenesis using Ag2S quantum dots in the NIR-II window

Improving the tissue penetration depth and spatial resolution of fluorescence-based optical nanoprobes remains a grand challenge for their practical applications in in vivo imaging, due to the scattering and absorption and endogenous autofluorescence of living tissues. Here, we present that Ag2S quantum dots (QDs), containing no toxic ions, exhibiting long circulation time and high stability, act as a new kind of fluorescent probes in the second near-infrared window (NIR-II, 1000–1350 nm) which enable in vivo monitoring of lymphatic drainage and vascular networks with deep tissue penetration and high spatial and temporal resolution. In addition, NIR-II fluorescence imaging with Ag2S QDs provide ultrahigh spatial resolution (∼40 μm) that permits us to track angiogenesis mediated by a tiny tumor (2–3 mm in diameter) in vivo. Our results indicate that Ag2S QDs are promising NIR-II fluorescent nanoprobes that could be useful in surgical treatments such as sentinel lymph node (SLN) dissection as well in assessment of blood supply in tissues and organs and screening of anti-angiogenic drugs.

Kite aerial photography for low-cost, ultra-high spatial resolution multi-spectral mapping of intertidal landscapes.

Intertidal ecosystems have primarily been studied using field-based sampling; remote sensing offers the ability to collect data over large areas in a snapshot of time that could complement field-based sampling methods by extrapolating them into the wider spatial and temporal context. Conventional remote sensing tools (such as satellite and aircraft imaging) provide data at limited spatial and temporal resolutions and relatively high costs for small-scale environmental science and ecologically-focussed studies. In this paper, we describe a low-cost, kite-based imaging system and photogrammetric/mapping procedure that was developed for constructing high-resolution, three-dimensional, multi-spectral terrain models of intertidal rocky shores. The processing procedure uses automatic image feature detection and matching, structure-from-motion and photo-textured terrain surface reconstruction algorithms that require minimal human input and only a small number of ground control points and allow the use of cheap, consumer-grade digital cameras. The resulting maps combine imagery at visible and near-infrared wavelengths and topographic information at sub-centimeter resolutions over an intertidal shoreline 200 m long, thus enabling spatial properties of the intertidal environment to be determined across a hierarchy of spatial scales. Results of the system are presented for an intertidal rocky shore at Jervis Bay, New South Wales, Australia. Potential uses of this technique include mapping of plant (micro- and macro-algae) and animal (e.g. gastropods) assemblages at multiple spatial and temporal scales.

Energy-resolved magnetic domain imaging in TbCo alloys by valence band photoemission magnetic circular dichroism

We report magnetic domain imaging of a terbium cobalt (TbCo) alloy thin film with a high perpendicular magnetic anisotropy in one- and two-photon photoemission electron microscopy (PEEM). Both photoemission processes deliver a clear magnetic circular dichroism (MCD) whose strength is strongly energy dependent. Comparing the energy dependence of the MCD signal in one- and two-photon photoemission, we conclude that the magnetic contrast is mainly an initial state effect. Our results ultimately show that MCD contrast can be obtained in PEEM in valence band photoemission from a material supporting all-optical magnetization switching. This opens the way for the investigation of the all-optical switching process with simultaneous ultrahigh temporal and spatial resolution.

ANTI-PARALLEL EUV FLOWS OBSERVED ALONG ACTIVE REGION FILAMENT THREADS WITH HI-C

Plasma flows within prominences/filaments have been observed for many years and hold valuable clues concerning the mass and energy balance within these structures. Previous observations of these flows primarily come from H-alpha and cool EUV lines (e.g., 304A) where estimates of the size of the prominence threads has been limited by the resolution of the available instrumentation. Evidence of `counter-steaming' flows has previously been inferred from these cool plasma observations but now, for the first time, these flows have been directly imaged along fundamental filament threads within the million degree corona (at 193A). In this work we present observations of an active region filament observed with Hi-C that exhibits anti-parallel flows along adjacent filament threads. Complementary data from SDO/AIA and HMI are presented. The ultra-high spatial and temporal resolution of Hi-C allow the anti-parallel flow velocities to be measured (70-80 km/s) and gives an indication of the resolvable thickness of the individual strands (0.8'' +/- 0.1''). The temperature distribution of the plasma flows was estimated to be log T(K) = 5.45 +/- 0.10 using EM loci analysis. We find that SDO/AIA cannot clearly observe these anti-parallel flows nor measure their velocity or thread width due to its larger pixel size. We suggest that anti-parallel/counter-streaming flows are likely commonplace within all filaments and are currently not observed in EUV due to current instrument spatial resolution.

Ultra-high resolution imaging of the human brain using acquisition-weighted imaging at 9.4 T

One of the main goals of ultra-high field MRI is to increase the spatial resolution reached in structural and functional images. Here, the possibility to obtain in vivo images of the human brain with voxel volumes below 0.02mm3 is shown at 9.4T.To optimize SNR and suppress ringing artifacts, an acquisition-weighted 3D gradient-echo sequence is used, which acquires more averages in the center than in the outer regions of k-space. The weighting function is adjusted to avoid losses in spatial resolution and scan duration compared to a conventional experiment with an equal number of scans and otherwise identical parameters. Spatial resolution and SNR of the weighted sequence are compared to conventionally acquired images by means of phantom and in vivo measurements, and show improved image quality with unchanged spatial resolution and an SNR increase of up to 36% in phantoms and 20%±5% in vivo. Ultra-high resolution images with a voxel volume of 0.014mm3 (0.13×0.13×0.8mm3) from the human brain have sufficient SNR and show fine intracortical detail, demonstrating the potential of the technique.The combination of acquisition-weighted imaging and highly sensitive array coils at ultra-high fields thus makes it possible to obtain images with ultra-high spatial resolutions within acceptable scan times.

Using an Unmanned Aerial Vehicle (UAV) to capture micro-topography of Antarctic moss beds

Mosses, the dominant flora of East Antarctica, show evidence of drying in recent decades, likely due to the regional effects of climate change. Given the relatively small area that such moss beds occupy, new tools are needed to map and monitor these fragile ecosystems in sufficient detail. In this study, we collected low altitude aerial photography with a small multi-rotor Unmanned Aerial Vehicle (UAV). Structure from Motion (SfM) computer vision techniques were applied to derive ultra-high resolution 3D models from multi-view aerial photography. A 2cm digital surface model (DSM) and 1cm orthophoto mosaic were derived from the 3D model and aerial photographs, respectively. The geometric accuracy of the orthophoto and DSM was 4cm. A weighted contributing upstream area was derived with the D-infinity algorithm, based on the DSM and a snow cover map derived from the orthophoto. The contributing upstream area was used as a proxy for water availability from snowmelt, one of the key environmental drivers of moss health. A Monte Carlo simulation with 300 realisations was implemented to model the impact of error in the DSM on runoff direction. Significant correlations were found between these simulated water availability values and field measurements of moss health and water content. In the future ultra-high spatial resolution DSMs acquired with a UAV could thus be used to determine the impact of changing snow cover on the health and spatial distribution of polar vegetation non-destructively.

On the problems of the application of atomic-force microscopes for studying the surface roughness of elements for imaging optics

The key problems of applying atomic-force microscopes when studying the surface roughness of elements for imaging optics are discussed. Special attention is paid to optics of ultrahigh spatial resolution with an operating wavelength of no shorter than 100 nm. It is proposed that a specialized microscope for optical applications equipped with a tetraaxial goniometer be developed.

Ultra-high resolution optical CT dosimetry for the visualisation of synchrotron microbeam therapy doses

Optical CT is a method that can potentially provide both accurate dosimetry at high spatial resolution and 3-D visualisation over a large field-of-view in a single dataset. The major factors limiting spatial resolution in previous studies are analysed here and it is shown that improvements in equipment specification can overcome many of these. The need for ultra-high spatial resolution in the verification of microbeam radiation therapy verification is demonstrated and example images of a PRESAGE® sample are presented.

Spatial resolution improvement and dose reduction potential for inner ear CT imaging using a z‐axis deconvolution technique

To assess the z-axis resolution improvement and dose reduction potential achieved using a z-axis deconvolution technique with iterative reconstruction (IR) relative to filtered backprojection (FBP) images created with the use of a z-axis comb filter. Each of three phantoms were scanned with two different acquisition modes: (1) an ultrahigh resolution (UHR) scan mode that uses a comb filter in the fan angle direction to increase in-plane spatial resolution and (2) a z-axis ultrahigh spatial resolution (zUHR) scan mode that uses comb filters in both the fan and cone angle directions to improve both in-plane and z-axis spatial resolution. All other scanning parameters were identical. First, the ACR CT Accreditation phantom, rotated by 90° so that the high-contrast spatial resolution targets were parallel to the coronal plane, was scanned to assess limiting spatial resolution and image noise. Second, section sensitivity profiles (SSPs) were measured using a copper foil embedded in an acrylic cylinder and the full-width-at-half-maximum (FWHM) and full-width-at-tenth-maximum (FWTM) of the SSPs were calculated. Third, an anthropomorphic head phantom containing a human skull was scanned to assess clinical acceptability for imaging of the temporal bone. For each scan, FBP images were reconstructed for the zUHR scan using the narrowest image thickness available. For the CT accreditation phantom, zUHR images were also reconstructed using an IR algorithm (SAFIRE, Siemens Healthcare, Forchheim, Germany) to assess the influence of the IR algorithm on image noise. A z-axis deconvolution technique combined with the IR algorithm was used to reconstruct images at the narrowest image thickness possible from the UHR scan data. Images of the ACR and head phantoms were reformatted into the coronal plane. The head phantom images were evaluated by a neuroradiologist to assess acceptability for use in patients undergoing clinically indicated CT imaging of the temporal bone. The limiting spatial resolution was 12 lp/cm for the FBP-zUHR images and the IR-UHR images, although visual assessment indicated a slight improvement for the IR-UHR images. Image noise was 213.0, 181.8, and 153.5 for the FBP-zUHR, IR-zUHR, and IR-UHR images, respectively. While the FWHM was essentially the same for the FBP-zUHR and IR-UHR images, the FWTM of the IR-UHR images was almost 50% smaller compared to the FBP-zUHR images (0.83 vs 1.25 mm, respectively). Images of the anthropomorphic head phantom were judged to be of higher quality for the IR-UHR images compared to the FBP-zUHR images. With use of a z-axis deconvolution technique, z-axis spatial resolution was improved for scans acquired using a comb filter only in the fan angle direction relative to FBP images acquired with a comb filter in both the fan and cone angle directions. By avoiding use of the comb filter in the cone angle direction and use of an IR algorithm, image noise was substantially reduced for the same scanner output (CTDIvol). Thus, overall image quality (spatial resolution and image noise) can be maintained relative to the FBP-zUHR technique at a lower radiation dose.

High‐harmonic generation with plasmonics: feasible or unphysical?

High-harmonic generation (HHG) provides for efficient and coherent XUV generation for a wide range of spectroscopic applications, enabling attosecond spectroscopy, ultrafast photo-emission spectroscopy, or ultrahigh spatial resolution imaging [1]. Conventionally HHG is based on the use of high power amplified femtosecond laser pulses, special phase matching techniques, or intra-cavity field enhancement. As a compelling alternative, resonant plasmonic field enhancement in metallic nanostructures has been proposed to achieve the necessary peak intensity for HHG. Using nJ pulses from a Ti:S oscillator, XUV generation attributed to plasmon-enhanced HHG was reported in 2008 by the group of Seung-Woo Kim from KAIST in Daejeon, Korea, first in Ar [2] and later

Quantitative x-ray phase imaging at the nanoscale by multilayer Laue lenses

For scanning x-ray microscopy, many attempts have been made to image the phase contrast based on a concept of the beam being deflected by a specimen, the so-called differential phase contrast imaging (DPC). Despite the successful demonstration in a number of representative cases at moderate spatial resolutions, these methods suffer from various limitations that preclude applications of DPC for ultra-high spatial resolution imaging, where the emerging wave field from the focusing optic tends to be significantly more complicated. In this work, we propose a highly robust and generic approach based on a Fourier-shift fitting process and demonstrate quantitative phase imaging of a solid oxide fuel cell (SOFC) anode by multilayer Laue lenses (MLLs). The high sensitivity of the phase to structural and compositional variations makes our technique extremely powerful in correlating the electrode performance with its buried nanoscale interfacial structures that may be invisible to the absorption and fluorescence contrasts.

Oral cancer diagnostics based on infrared spectral markers and wax physisorption kinetics

Infrared microspectroscopy is an emerging approach for disease analysis owing to its capability for in situ chemical characterization of pathological processes. Synchrotron-based infrared microspectroscopy (SR-IMS) provides ultra-high spatial resolution for profiling biochemical events associated with disease progression. Spectral alterations were observed in cultured oral cells derived from healthy, precancerous, primary, and metastatic cancers. An innovative wax-physisorption-based kinetic FTIR imaging method for the detection of oral precancer and cancer was demonstrated successfully. The approach is based on determining the residual amount of paraffin wax (C(25)H(52)) or beeswax (C(46)H(92)O(2)) on a sample surface after xylene washing. This amount is used as a signpost of the degree of physisorption that altered during malignant transformation. The results of linear discriminant analysis (LDA) of oral cell lines indicated that the methylene (CH(2)) and methyl group (CH(3)) stretching vibrations in the range of 3,000-2,800 cm(-1) have the highest accuracy rate (89.6 %) to discriminate the healthy keratinocytes (NHOK) from cancer cells. The results of wax-physisorption-based FTIR imaging showed a stronger physisorption with beeswax in oral precancerous and cancer cells as compared with that of NHOK, which showed a strong capability with paraffin wax. The infrared kinetic study of oral cavity tissue showed a consistency in the wax physisorption of the cell lines. On the basis of our findings, these results show the potential use of wax-physisorption-based kinetic FTIR imaging for the early screening of oral cancer lesions and the chemical changes during oral carcinogenesis.

Individual Deciduous Tree Recognition in Leaf-Off Aerial Ultrahigh Spatial Resolution Remotely Sensed Imagery

This study proposed and tested a multistep method for the recognition of individual deciduous trees in leaf-off aerial ultrahigh spatial resolution remotely sensed (UHSRRS) imagery. This topic has received limited coverage in previous endeavors, which focused mainly on the detection and delineation of coniferous trees in remotely sensed images with relatively lower spatial resolutions. Thus, the traditional algorithms tend to fail in case of the referred scenario. In order to fill this technical gap, an algorithm that joins mathematical morphological operations and marker-controlled watershed segmentation was first assumed for the extraction of single trees in UHSRRS images. Next, a distribution-free support vector machine (SVM) classifier was applied to distinguish the extracted segments as deciduous or coniferous trees, merely in terms of two newly-derived morphological features. Experimental evaluations indicated that the integral solution plan can extract and classify the deciduous and coniferous trees in the leaf-off aerial UHSRRS images of local dense forest for test with correctness over 92% and 70%, respectively. Overall, the recognition results with >;66% correctness have primarily validated the proposed technique.

A Line Pair Indicator Made of Gd Film for Evaluating Spatial Resolution

A device for evaluating the spatial resolution of a neutron imaging system was developed. Using laser processes, line- pair patterns were fabricated on a 0.005-mm-thick Gd film evaporated on a glass plate. Large line pairs of widths ranging from 0.2 to 2 mm were machined using an Nd:YVO 4 laser (0.03 mm spot size) and displayed on a brightness field, while small line pairs of widths ranging from 0.01 to 0.1 mm were created using an eximer laser on a dark field. A scanning electron microscope (SEM) observation determined that, although the large line pairs machined using the Nd:YVO 4 laser were chipped on the corners of the Gd bars, the difference between the measured and designed line- pair widths was controlled absolutely within 0.02 mm. In the small line pairs of less than 0.1 mm width processed using the eximer laser, edges sharper than those of the large line pairs were formed. In neutron imaging tests using a LiF/ZnS(Ag) scintillator and a CCD camera system, good contrast images were obtained with the brightness field, even at 0.005 mm thickness. The small line pairs on the dark field were observed using a LiF single crystal detector having an ultra-high spatial resolution of approximately 0.005 mm. Splits in the small line pairs of as little as 0.01 mm wide were shown with good contrast on the images.

Integration of ultra-high field MRI and histology for connectome based research of brain disorders

Ultra-high field magnetic resonance imaging (MRI) became increasingly relevant for in vivo neuroscientific research because of improved spatial resolutions. However, this is still the unchallenged domain of histological studies, which long played an important role in the investigation of neuropsychiatric disorders. While the field of biological psychiatry strongly advanced on macroscopic levels, current developments are rediscovering the richness of immunohistological information when attempting a multi-level systematic approach to brain function and dysfunction. For most studies, histology sections lost information on three-dimensional reconstructions. Translating histological sections to 3D-volumes would thus not only allow for multi-stain and multi-subject alignment in post mortem data, but also provide a crucial step in big data initiatives involving the network analyses currently performed with in vivo MRI. We therefore investigated potential pitfalls during integration of MR and histological information where no additional blockface information is available. We demonstrated that strengths and requirements from both methods can be effectively combined at a spatial resolution of 200 μm. However, the success of this approach is heavily dependent on choices of hardware, sequence and reconstruction. We provide a fully automated pipeline that optimizes histological 3D reconstructions, providing a potentially powerful solution not only for primary human post mortem research institutions in neuropsychiatric research, but also to help alleviate the massive workloads in neuroanatomical atlas initiatives. We further demonstrate (for the first time) the feasibility and quality of ultra-high spatial resolution (150 μm isotopic) imaging of the entire human brain MRI at 7T, offering new opportunities for analyses on MR-derived information.

Conditional Tat protein expression in the GT-tg bigenic mouse brain induces gray matter density reductions

Tat (Trans-activator of transcription) is implicated in the neuropathogenesis of HIV-1 infection and known to contribute to neuronal damage and learning and memory impairments. However, direct neuroanatomical demonstration of Tat pathobiology is limited. GT-tg bigenic mice with a doxycycline (Dox)-inducible and brain-selective tat gene were used to test the hypothesis that conditional induction of Tat activity in brain can induce gray matter density abnormalities. Ultra high spatial resolution ex vivo magnetic resonance imaging (MRI) combined with a voxel based morphometry (VBM) analysis revealed gray matter density reductions in the sublenticular extended amygdala, the amygdala, the amygdala-hippocampal area, piriform and peri-/entorhinal cortices, and hypothalamus, in Tat-expressing GT-tg mice compared to Dox-treated C57Bl/6J mice. These neuroanatomical abnormalities are consistent with regions expected to be abnormal based on behavioral deficits exhibited by Tat-expressing mice (Carey et al., 2012). These experiments provide the first neuroimaging evidence that conditional Tat protein expression in the GT-tg bigenic mouse model alters brain structure. The findings warrant future studies to further characterize effects of conditional Tat expression on brain structure. Such studies may improve our understanding of the neurological underpinnings of neuroAIDS and the neurodegeneration associated with HIV-1 infection, potentially leading to new treatments.

High and Ultra-High resolution processing of satellite Sea Surface Temperature data over Southern European Seas in the framework of MyOcean project

In the framework of the GMES (Global Monitoring for Environment and Security) MyOcean projects, funded by the European Commission, different remotely-sensed Sea Surface Temperature (SST) supercollated (L3S) and interpolated (L4) data are produced and distributed in near-real time by the Consiglio Nazionale delle Ricerche – Istituto di Scienze dell'Atmosfera e del Clima – Gruppo di Oceanografia da Satellite (CNR). These SST products are based on the night-time images collected by the infrared sensors mounted on different satellite platforms and cover the whole Southern European Seas (i.e. the Mediterranean Sea, including the eastern Atlantic Ocean, and the Black Sea). The CNR processing chains include several modules, from the data extraction and preliminary quality control, to cloudy pixel removal and satellite images collating/merging up to L3S. A two-step algorithm finally allows to interpolate SST data at High (HR 1/16°) and Ultra-High (UHR 1/100°) spatial resolution. The basic design and the main algorithms used in MyOcean processing chains are described hereafter. A validation of the HR super-collating procedure developed during MyOcean is presented. The Mediterranean HR and UHR L3S and L4 operational products have then been validated vs in situ SST measurements from drifting buoys, covering two years of data (2010–2011). The validation includes a comparison between the UHR CNR L4 and the G1SST L4 developed by Chao et al. (2009).

On‐Demand Patterning of Nanostructured Pentacene Transistors by Scanning Thermal Lithography

Thermal scanning lithography is used to pattern semiconducting nanoribbon-like pentacene structures with ultrahigh spatial resolution onto arbitrary substrates in air. The method allows control of the pentacene crystal growth direction and domain-size distribution. By combining these quasi-one-dimensional nanoribbon-like structures with conductive electrodes and a suitable gate dielectric, functional p-channel transistors are demonstrated.

Ultra high spatial and temporal resolution breast imaging at 7T

There is a need to obtain higher specificity in the detection of breast lesions using MRI. To address this need, Dynamic Contrast-Enhanced (DCE) MRI has been combined with other structural and functional MRI techniques. Unfortunately, owing to time constraints structural images at ultra-high spatial resolution can generally not be obtained during contrast uptake, whereas the relatively low spatial resolution of functional imaging (e.g. diffusion and perfusion) limits the detection of small lesions. To be able to increase spatial as well as temporal resolution simultaneously, the sensitivity of MR detection needs to increase as well as the ability to effectively accelerate the acquisition. The required gain in signal-to-noise ratio (SNR) can be obtained at 7T, whereas acceleration can be obtained with high-density receiver coil arrays. In this case, morphological imaging can be merged with DCE-MRI, and other functional techniques can be obtained at higher spatial resolution, and with less distortion [e.g. Diffusion Weighted Imaging (DWI)]. To test the feasibility of this concept, we developed a unilateral breast coil for 7T. It comprises a volume optimized dual-channel transmit coil combined with a 30-channel receive array coil. The high density of small coil elements enabled efficient acceleration in any direction to acquire ultra high spatial resolution MRI of close to 0.6 mm isotropic detail within a temporal resolution of 69 s, high spatial resolution MRI of 1.5 mm isotropic within an ultra high temporal resolution of 6.7 s and low distortion DWI at 7T, all validated in phantoms, healthy volunteers and a patient with a lesion in the right breast classified as Breast Imaging Reporting and Data System (BI-RADS) IV.

Detection of Single Influenza Viral RNA in Cells Using a Polymeric Sequence Probe

A polynucleotide probe, call a polymeric sequence probe (PSP), was used to detect influenza A (Influenza A/WSN/33) NA (Neuraminidase) viral RNA in Madin-Darby canine kidney (MDCK) cells. The PSP is a single-stranded DNA molecule with ~2,000 tandem repeat fluorescence binding sites and target binding sites that can bind with multiple fluorescence complementary oligos and target viral RNA using a fluorescence in situ hybridization (FISH) process. A single viral RNA labeled by PSP can be directly observed in MDCK cells. The simple FISH protocol enables the observation and quantitative analysis of the infectious process and drug effects with ultrahigh sensitivity and spatial resolution.