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PubMed2000

Reconstruction of axial tomographic high resolution data from confocal fluorescence microscopy: a method for improving 3D FISH images.

Fluorescent confocal laser scanning microscopy allows an improved imaging of microscopic objects in three dimensions. However, the resolution along the axial direction is three times worse than the resolution in lateral directions. A method to overcome this axial limitation is tilting the object under the microscope, in a way that the direction of the optical axis points into different directions relative to the sample. A new technique for a simultaneous reconstruction from a number of such axial tomographic confocal data sets was developed and used for high resolution reconstruction of 3D-data both from experimental and virtual microscopic data sets. The reconstructed images have a highly improved 3D resolution, which is comparable to the lateral resolution of a single deconvolved data set. Axial tomographic imaging in combination with simultaneous data reconstruction also opens the possibility for a more precise quantification of 3D data. The color images of this publication can be accessed from http://www.esacp.org/acp/2000/20-1/heintzmann.++ +htm. At this web address an interactive 3D viewer is additionally provided for browsing the 3D data. This java applet displays three orthogonal slices of the data set which are dynamically updated by user mouse clicks or keystrokes.

Algorithms↗

PubMed1997

Colour bit-stealing to enhance the luminance resolution of digital displays on a single pixel basis.

A look-up table algorithm is described for enhancing the luminance resolution at the expense of the colour resolution in digital displays of colour images. For colour displays with a look-up table resolution of 8 bits/gun, the algorithm provides a luminance resolution of 11-12 bits without loss of spatial or temporal resolution. This improvement can reduce the minimum luminance step in the mid-luminance range from 1.5% to > 0.2% with no additional hardware.

Algorithms↗

PubMed2006

A gene-based high-resolution comparative radiation hybrid map as a framework for genome sequence assembly of a bovine chromosome 6 region associated with QTL for growth, body composition, and milk performance traits.

BACKGROUND: A number of different quantitative trait loci (QTL) for various phenotypic traits, including milk production, functional, and conformation traits in dairy cattle as well as growth and body composition traits in meat cattle, have been mapped consistently in the middle region of bovine chromosome 6 (BTA6). Dense genetic and physical maps and, ultimately, a fully annotated genome sequence as well as their mutual connections are required to efficiently identify genes and gene variants responsible for genetic variation of phenotypic traits. A comprehensive high-resolution gene-rich map linking densely spaced bovine markers and genes to the annotated human genome sequence is required as a framework to facilitate this approach for the region on BTA6 carrying the QTL. RESULTS: Therefore, we constructed a high-resolution radiation hybrid (RH) map for the QTL containing chromosomal region of BTA6. This new RH map with a total of 234 loci including 115 genes and ESTs displays a substantial increase in loci density compared to existing physical BTA6 maps. Screening the available bovine genome sequence resources, a total of 73 loci could be assigned to sequence contigs, which were already identified as specific for BTA6. For 43 loci, corresponding sequence contigs, which were not yet placed on the bovine genome assembly, were identified. In addition, the improved potential of this high-resolution RH map for BTA6 with respect to comparative mapping was demonstrated. Mapping a large number of genes on BTA6 and cross-referencing them with map locations in corresponding syntenic multi-species chromosome segments (human, mouse, rat, dog, chicken) achieved a refined accurate alignment of conserved segments and evolutionary breakpoints across the species included. CONCLUSION: The gene-anchored high-resolution RH map (1 locus/300 kb) for the targeted region of BTA6 presented here will provide a valuable platform to guide high-quality assembling and annotation of the currently existing bovine genome sequence draft to establish the final architecture of BTA6. Hence, a sequence-based map will provide a key resource to facilitate prospective continued efforts for the selection and validation of relevant positional and functional candidates underlying QTL for milk production and growth-related traits mapped on BTA6 and on similar chromosomal regions from evolutionary closely related species like sheep and goat. Furthermore, the high-resolution sequence-referenced BTA6 map will enable precise identification of multi-species conserved chromosome segments and evolutionary breakpoints in mammalian phylogenetic studies.

Animals↗

PubMed2005

A streamlined method for systematic, high resolution in situ analysis of mRNA distribution in plants.

BACKGROUND: In situ hybridisation can provide cellular, and in some cases sub-cellular, resolution of mRNA levels within multicellular organisms and is widely used to provide spatial and temporal information on gene expression. However, standard protocols are complex and laborious to implement, restricting analysis to one or a few genes at any one time. Whole-mount and reverse transcriptase-PCR (RT-PCR) based protocols increase throughput, but can compromise both specificity and resolution. With the advent of genome-wide analysis of gene expression, there is an urgent need to develop high-throughput in situ methods that also provide high resolution. RESULTS: Here we describe the development of a method for performing high-throughput in situ hybridisations that retains both the high resolution and the specificity of the best manual versions. This refined semi-automated protocol has the potential for determining the spatial and temporal expression patterns of hundreds of genes in parallel on a variety of tissues. We show how tissue sections can be organized on microscope slides in a manner that allows the screening of multiple probes on each slide. Slide handling, hybridisation and processing steps have been streamlined providing a capacity of at least 200 probes per week (depending on the tissue type). The technique can be applied easily to different species and tissue types, and we illustrate this with wheat seed and Arabidopsis floral meristems, siliques and seedlings. CONCLUSION: The approach has the high specificity and high resolution of previous in situ methods while allowing for the analysis of several genes expression patterns in parallel. This method has the potential to provide an analysis of gene expression patterns at the genome level.

Journal Article↗

PubMed2005

High-resolution optical coherence tomographic imaging of osteoarthritic cartilage during open knee surgery.

This study demonstrates the first real-time imaging in vivo of human cartilage in normal and osteoarthritic knee joints at a resolution of micrometers, using optical coherence tomography (OCT). This recently developed high-resolution imaging technology is analogous to B-mode ultrasound except that it uses infrared light rather than sound. Real-time imaging with 11-microm resolution at four frames per second was performed on six patients using a portable OCT system with a handheld imaging probe during open knee surgery. Tissue registration was achieved by marking sites before imaging, and then histologic processing was performed. Structural changes including cartilage thinning, fissures, and fibrillations were observed at a resolution substantially higher than is achieved with any current clinical imaging technology. The structural features detected with OCT were evident in the corresponding histology. In addition to changes in architectural morphology, changes in the birefringent or the polarization properties of the articular cartilage were observed with OCT, suggesting collagen disorganization, an early indicator of osteoarthritis. Furthermore, this study supports the hypothesis that polarization-sensitive OCT may allow osteoarthritis to be diagnosed before cartilage thinning. This study illustrates that OCT, which can eventually be developed for use in offices or through an arthroscope, has considerable potential for assessing early osteoarthritic cartilage and monitoring therapeutic effects for cartilage repair with resolution in real time on a scale of micrometers.

Aged↗

PubMed1994

The limit of tactile spatial resolution in humans: grating orientation discrimination at the lip, tongue, and finger.

The sensory neural pathways serving the lip, tongue, and finger are specialized for spatial information processing; thus, damage to these pathways is likely to be manifested most prominently as a loss of spatial acuity. For that reason, accurate measurement of spatial resolution at these regions is particularly important. The conventional test, the two-point discrimination task, does not measure the limit of spatial resolution and it yields variable results because it does not control nonspatial cues. The aim of this study was to quantify the limits of spatial resolution at the lip, tongue, and finger and to study the repeatability of those measurements using a stimulus that does not introduce nonspatial cues. We employed a grating orientation discrimination test, which has been studied extensively in relation to the underlying neural mechanisms. We obtained psychophysical thresholds for tactile spatial resolution from 15 normal, young adult subjects over seven test sessions. The finest gratings whose orientations were discriminated reliably had groove widths (gratings had equal groove and bar widths) that averaged 0.51 mm at the lip, 0.58 mm at the tongue, and 0.94 mm at the finger. These threshold measurements were highly reproducible between sessions with an overall improvement of 2% per session. These data suggest that the grating orientation discrimination task provides a stable, reliable measure of the human capacity for spatial resolution.

Adult↗

PubMed2003

Rate of resolution of histologically verified intracranial tuberculomas.

OBJECTIVE: The goal of this study was to determine the rate of radiological resolution of histopathologically proven tuberculomas treated with antituberculous therapy (ATT). The effects of the size of the tuberculomas, the number of tuberculomas, and the addition of corticosteroid therapy on the rate of resolution of the tuberculomas were also studied. METHODS: Twenty-eight patients (age range, 5-48 yr; 14 male and 14 female patients) with histologically proven intracranial tuberculomas were prospectively monitored with contrast-enhanced computed tomographic scans. The patients received ATT consisting of rifampicin and isoniazid for a period of 18 months, with ethambutol and/or pyrazinamide for a minimum of 3 months. Fifteen patients also received corticosteroid therapy for 1 to 6 weeks. Of the 28 patients, 17 patients underwent partial excision, 6 underwent open biopsy, and 5 underwent stereotactic biopsy of their tuberculomas. RESULTS: Kaplan-Meier analysis revealed that, after 9 months of ATT, only 18.2% of the patients demonstrated complete resolution of their tuberculomas; even after 18 months of ATT, 69.2% of the patients had residual lesions. By 24 months, 54% of the patients demonstrated complete resolution of their tuberculomas. Although the number of tuberculomas, corticosteroid administration, prior treatment with ATT, and the duration of symptoms before presentation (<6 mo versus >6 mo) did not influence the rate of resolution, larger tuberculomas (maximal size, >4 cm) were observed to resolve more slowly than smaller tuberculomas (<4 cm) (P = 0.02). CONCLUSION: More than two-thirds of patients with partially excised or biopsied intracranial tuberculomas exhibited persistent lesions on computed tomographic scans, even after 18 months of ATT. Therefore, the duration of ATT for patients with intracranial tuberculomas should be based on the radiological responses of the tuberculomas. Our data suggest that some patients with intracranial tuberculomas might require prolonged periods of ATT.

Adolescent↗

PubMed2003

Visual resolution of gratings by the compound eye of the bee Apis mellifera.

Early measurements of the resolution of horizontal versus vertical gratings were confirmed, with a limit near a period of 2.5 degrees, and the resolution is similar when vertical or horizontal gratings are tested separately against grey. Bees were next trained to discriminate from a distance between gratings at 45 degrees versus 135 degrees, with no green contrast, on targets presented in a vertical plane at a fixed distance. As expected, they fail to learn; however, with green contrast but no modulation difference the resolution limit is near 3.5 degrees. With vertical and horizontal gratings with no green contrast they discriminate but do not learn an orientation cue. In order to eliminate the orientation cue altogether, new bees were then trained with alternating vertical and horizontal gratings versus grey, or with a black and white checkerboard versus grey. Tests of these trained bees with horizontal or with vertical gratings separately against grey again show a resolution down to a period near 2.5 degrees. These results, taken together, show that when edge orientation alone is the cue, the limit of resolution is near 3.5 degrees, but when receptor modulation is the cue, the limit is near 2.5 degrees.

Animals↗

PubMed1996

The variation of resolution and of ommatidial dimensions in the compound eyes of the fiddler crab Uca lactea annulipes (Ocypodidae, Brachyura, Decapoda)

We studied variations in the optical properties of the compound eyes of Uca lactea annulipes using in vivo optical and histological techniques. The distribution of resolving power in the eyes of this fiddler crab species is typical for arthropods that inhabit flat environments: the eyes possess a panoramic equatorial acute zone for vertical resolution and a steep decrease of resolution away from the eye equator in the dorsal and ventral visual fields. The dimensions of the cellular components of the ommatidia vary accordingly: in the equatorial part of the eyes, facets are larger, and crystalline cones and rhabdoms are longer than in the dorsal and ventral parts of the eyes. Along the eye equator, horizontal resolution is low compared with vertical resolution and varies little throughout the visual field. The eyes of Uca lactea annulipes are unusual in that the gradient of vertical anatomical and optical resolution is steeper in the dorsal than in the ventral visual field. We interpret this difference as indicating that the information content of the world as seen by the crabs differs above and below the horizon line in specific and predictable ways.

Journal Article↗

PubMed1983

The spatial resolution of a rotating gamma camera tomographic facility.

An important feature determining the spatial resolution in transverse sections reconstructed by convolution and back-projection is the frequency filter corresponding to the convolution kernel. Equations have been derived giving the theoretical spatial resolution, for a perfect detector and noise-free data, using four filter functions. Experiments have shown that physical constraints will always limit the resolution that can be achieved with a given system. The experiments indicate that the region of the frequency spectrum between KN/2 and KN where KN is the Nyquist frequency does not contribute significantly to resolution. In order to investigate the physical effect of these filter functions, the spatial resolution of reconstructed images obtained with a GE 400T rotating gamma camera has been measured. The results obtained serve as an aid to choosing appropriate reconstruction filters for use with a rotating gamma camera system.

Air↗

PubMed1989

Radiological evaluation of temporal bone disease: high-resolution computed tomography versus conventional X-ray diagnosis.

Sixty-two patients with different temporal bone lesions were prospectively examined by high-resolution computed tomography (CT) and conventional plain radiography, including pluridirectional tomography. High-resolution CT enabled a clear diagnosis in 80% of cases, conventional radiology in 63%; 1.6-times more bone information was recorded by high-resolution CT which is clearly superior for imaging cholesteatomas, metastases and inflammatory processes and for evaluating osseous destruction. With regard to pathological soft tissue or effusions filling the tympanic cavities, conventional radiology shows poor sensitivity (0.61). High-resolution CT is the most sensitive method for the imaging and classification of temporal bone fractures, including labyrinthine damage and ossicular chain injuries. Only in cases of atypical fractures with an unfavourable relationship to the CT planes, can carefully directed tomography be more effective. In most cases high-resolution CT replaces conventional radiology and should be the method of choice for comprehensive radiological examination of the temporal bone.

Bone Diseases↗

PubMed1994

Bitewing-based alveolar bone densitometry: digital imaging resolution requirements.

The resolution requirements were determined for detection of incremental alveolar bone loss from digitized Ektaspeed radiographs. Ten clinical radiographs were examined with a calibrated optical microscope to measure the smallest feature of interest discernible for alveolar bone. Images of trabeculae > 100 microns in diameter could be identified, but smaller ones could not be resolved. The Nyquist Criterion requires sampling with 50 microns (or smaller) pixels to measure such features. Numerous 25 microns Ag aggregates were present. Fifty microns resolution is a practical compromise between noise level and feature resolution. In another example of 10 bitewings digitized at 8 bit grey scale, about half the available range was used for alveolar bone, resulting in uncertainty, over a range of 2 optical density (OD) units, of about 1.42% at the average OD for alveolar bone (1.1). With the same radiographs digitized at 12 bit resolution, over 2000 of 4096 grey scales were used with a corresponding uncertainty of about 0.09%. Twelve-bit precision was also able to resolve smaller increments in an aluminium stepwedge than was 8 bit precision. Twelve-bit grey scale and 50 microns spatial resolution are recommended for alveolar bone densitometry performed with Ektaspeed radiographs.

Absorptiometry, Photon↗

PubMed2002

Characterization of a CCD-based digital x-ray imaging system for small-animal studies: properties of spatial resolution.

A digital x-ray imaging system was designed for small-animal studies. This system is a fiber-optics taper-coupled imaging system with two CCD arrays uniquely jointed. The x-ray source of the system has a small focal spot of 20 microm. This digital imaging system contains specially designed shelves to provide magnification levels, ranging from 1.5x to 5x. The system is characterized in terms of its properties of spatial resolution. An observer-based spatial resolution measurement was conducted with a line-pair target and a sector test pattern. The modulation transfer function of the system, with different magnifications, was studied by use of a 10-microm lead slit. The average resolutions at 50% and 5% modulations at 1x magnification were measured as 3.9 and 8.4 lp/mm, respectively, where lp indicates line pairs. With 5x magnification, the 50% and the 5% modulations provided 13.2- and 29.9-lp/mm, respectively, average spatial resolutions. The measurements showed consistency between the two individual CCD arrays; the difference in resolution between the two CCDs is less than 1%, even at high magnifications.

Animals↗

PubMed2006

High-resolution ab initio three-dimensional x-ray diffraction microscopy.

Coherent x-ray diffraction microscopy is a method of imaging nonperiodic isolated objects at resolutions limited, in principle, by only the wavelength and largest scattering angles recorded. We demonstrate x-ray diffraction imaging with high resolution in all three dimensions, as determined by a quantitative analysis of the reconstructed volume images. These images are retrieved from the three-dimensional diffraction data using no a priori knowledge about the shape or composition of the object, which has never before been demonstrated on a nonperiodic object. We also construct two-dimensional images of thick objects with greatly increased depth of focus (without loss of transverse spatial resolution). These methods can be used to image biological and materials science samples at high resolution with x-ray undulator radiation and establishes the techniques to be used in atomic-resolution ultrafast imaging at x-ray free-electron laser sources.

Algorithms↗

PubMed1997

Pulmonary alveolar proteinosis: high-resolution CT, chest radiographic, and functional correlations.

STUDY OBJECTIVE: To determine whether a correlation exists between pulmonary function and both frontal chest radiographs and high-resolution chest CT findings in patients with pulmonary alveolar proteinosis (PAP). DESIGN: Retrospective review of radiographic and clinical data. SETTING: Tertiary referral hospital. PATIENTS: Seven patients with PAP were studied on 25 occasions using high-resolution chest CT (n=21), frontal chest radiographs (n=19), and pulmonary function tests (PFTs) (n=25). MEASUREMENTS AND RESULTS: Visual estimates of the extent, degree, and overall severity of parenchymal abnormalities were determined for plain radiographs and high-resolution chest CT, and were correlated with PFTs. With high-resolution CT, the extent and severity of ground-glass opacity correlated significantly with the presence of a restrictive ventilatory defect, reduced diffusing capacity, and hypoxemia. Chest radiographic findings also correlated significantly with restrictive ventilatory defect, diffusing capacity, and hypoxemia. CONCLUSION: In patients with PAP, although high-resolution CT correlates more closely with pulmonary function, plain radiographs should be sufficient for follow-up.

Adult↗

PubMed1999

[High-resolution magnetoencephalography--studies with a small-volume phantom].

To investigate the spatiotemporal organisation of neuronal processes in an animal model using magnetoencephalography (MEG), a high temporal resolution (ms) and an appropriate spatial resolution of about 1 mm is necessary. With the aim of determining the localization error and the resolution power of high-resolution MEG systems, we developed a phantom capable of simulating the characteristics of animal models. The phantom enables us to variably position at least two magnetic field sources to within 0.1 mm. For source localization on the basis of the magnetic field data, a spatial filtering algorithm was used. The investigation of a 16-channel micro SQUID-MEG system with a current dipole orientated tangentially to the phantom surface produced the following localization data (min ... max, x, y--horizontal plane, z--depth); systematic localization error e(x) = 1.16 ... 1.67 mm, e(y) = -1.01 ... -1.28 mm, e(z) = -5.22 ... -7.64 mm, standard deviation of the individual measurements perpendicular to the dipole axis s(perp) = 0.05 ... 0.22 mm, along this axis s(long) = 0.20 ... 1.73 mm, in the depths sz = 0.17 ... 3.17 mm. The "goodness of fit" was > 95%. Separation of two dipoles was still possible for parallel dipoles at a distance apart of d(parallel) = 0.03 mm and for those oriented perpendicularly to each other at a distance apart of d(perp) = 0.10 mm. On the basis of these results we conclude that the MEG system can achieve a resolution sufficient to permit the investigation of neuronal microstructures. The spatial errors detected were related to sensor position in the cryostatic vessel as well as to external low-frequency noise.

Magnetoencephalography↗

PubMed1999

Development of capillary electrophoresis as an alternative to high resolution agarose electrophoresis for the diagnosis of multiple sclerosis.

The presence of oligoclonal bands in cerebrospinal fluid (CSF) is used as a diagnostic indicator of multiple sclerosis (MS). These bands, gamma-globulins thought to result from a restricted antibody response directed against autoantigens or viral antigens, are consistent with CSF-specific immunoglobulin synthesis when observed in the spinal fluid and not in the serum. Current methodology commonly involves electrophoresing concentrated CSF with high-resolution agarose gel electrophoresis (HRAGE) followed by protein staining in order to visualize the oligoclonal bands. Capillary zone electrophoresis (CZE) was evaluated as an alternative method. Separation of CSF and serum proteins from 54 patients in a bare silica capillary containing a high pH borate buffer allowed for resolution of the five major zones including the gamma-region and showed a 90% concordance with the results from HRAGE banding studies. Since a simple borate buffer did not provide adequate resolution of the oligoclonal bands in the gamma-region, the separation buffer was augmented with polyethylene glycol (PEG) which provided a significant enhancement in resolution of proteins in this region (24 patient study). In addition to obtaining banding information from electropherograms obtained with these separation conditions, it was feasible to calculate a CSF Index which compared favorably with the results from nephelometry. Finally, we show that zwitterionic additives such as O-phosphorylethanolamine may further enhance resolution and that capallary electrophoresis (CE) may allow oligoclonal banding information to be obtained directly from CSF without concentration.

Buffers↗

PubMed2004

Can the low-resolution structures of photointermediates of bacteriorhodopsin explain their crystal structures?

To understand the molecular mechanism of light-driven proton pumps, the structures of the photointermediates of bacteriorhodopsin have been intensively investigated. Low-resolution diffraction techniques have demonstrated substantial conformational changes at the helix level in the M and N intermediates, between which there are noticeable differences. The intermediate structures at atomic resolution have also been solved by x-ray crystallography. Although the crystal structures have demonstrated local structural changes, such as hydrogen bond network rearrangements including water molecules, the large conformational changes at the helix level are not necessarily observed. Furthermore, the two reported crystal structures of an intermediate accumulated using a common method were distinct. To reconcile these apparent discrepancies, low-resolution projection maps were calculated from the crystal structures and compared to the low-resolution intermediate structures obtained using native membranes. The crystal structures can be categorized into three groups, which qualitatively correspond to the low-resolution structures of the M1-type, M2-type, and N-type determined in the native membrane. Based on these results, we conclude that at least three types of intermediate structures play a role during the photocycle.

Bacteriorhodopsins↗