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PubMed1999

Determination of the Effective Ground State Potential Energy Function of Ozone from High-Resolution Infrared Spectra.

The effective ground state potential energy function of the ozone molecule near the C(2v) equilibrium configuration was obtained in a least-squares fit to the largest sample of experimental, high-resolution vibration-rotation data used for this purpose so far. The fitting is based on variational calculations carried out with the extended Morse Oscillator Rigid Bender Internal Dynamics model. The potential function is expanded in Morse-type functions of the stretching variables and in cosine of the bending angle. The present calculation produces results in significantly better agreement with experiment than previous determinations of the potential energy surface, and the energies predicted with the new surface are sufficiently accurate to be useful for the assignment of new high-resolution spectra. The rms (root-mean-square) deviation of the fit of rovibrational data up to J = 5 is 0.02 cm(-1). For the set of all 60 band centers of the (16)O(3) molecule included in the Atlas of Ozone Line Parameters, the rms deviation is 0.025 cm(-1), and for all band centers determined so far from high-resolution spectra, including those recently observed and assigned in Reims corresponding to highly excited stretching and bending vibrations (v(1) + v(2) + v(3) = 6), the rms deviation is 0.1 cm(-1). The "dark states" that produce resonance perturbations in the observed bands are described with experimental accuracy up to the (v(1)v(2)v(3)) = (080) state. Extrapolation tests demonstrate the predictive power of the potential function obtained: rotational extrapolation up to J = 10 for the 11 lowest vibrational states results in an rms deviation of 0.06cm(-1). Also, vibrational energies measured by low-resolution Raman spectroscopy (which were not included in the input data for the fit) are calculated within the experimental accuracy (rms = 1.6 cm(-1)) of the experimental values up to the dissociation limit. The statistical analysis suggests that the accuracy of the equilibrium geometry and force constants of the molecule is considerably improved relative to previous determinations. The long-range behavior of the fitted potential at the dissociation limit O(3) --> O(2) + O shows very good agreement with experimental data. The new potential energy surface was used to predict the band centers of the isotopomers (17)O(3) and (18)O(3). Copyright 1999 Academic Press.

Journal Article↗

PubMed1997

Dual-axis tomography: an approach with alignment methods that preserve resolution.

Tomographic reconstructions of biological specimens are now routinely being generated in our high voltage electron microscope by tilting the specimen around two orthogonal axes. Separate tomograms are computed from each tilt series. The two tomograms are aligned to each other with general 3-D linear transformations that can correct for distortions between the two tomograms, thus preserving the inherent resolution of the reconstruction throughout its volume. The 3-D Fourier transforms of the two tomograms are then selectively combined to achieve a single tomogram. Unlike a single-axis tomogram, a dual-axis tomogram shows good resolution for extended features at any orientation in the plane of the specimen; it also has improved resolution in the depth of the specimen. Calculations indicate that the improvements available from double tilting and from tilting to higher angles are largely additive. Actual and model data were used to assess whether varying the increment between tilted views in proportion to the cosine of the tilt angle would allow a reduction in the number of pictures required to achieve a given resolution of reconstruction. Analysis by Fourier sector correlation indicated that the variable tilt increment improved the reconstruction in some respects but degraded it in others. A varying tilt increment thus does not give an unqualified improvement, at least when using back-projection algorithms for the reconstruction.

Animals↗

PubMed1998

4Pi-confocal microscopy provides three-dimensional images of the microtubule network with 100- to 150-nm resolution.

We show the applicability of 4Pi-confocal microscopy to three-dimensional imaging of the microtubule network in a fixed mouse fibroblast cell. Comparison with two-photon confocal resolution reveals a fourfold better axial resolution in the 4Pi-confocal case. By combining 4Pi-confocal microscopy with Richardson-Lucy image restoration a further resolution increase is achieved. Featuring a three-dimensional resolution in the range 100-150 nm, the 4Pi-confocal (restored) images are intrinsically more detailed than their confocal counterparts. Our images constitute what to our knowledge are the best-resolved three-dimensional images of entangled cellular microtubules obtained with light to date.

3T3 Cells↗

PubMed1999

Review: resolution issues in single-particle reconstruction.

Specific factors that affect the resolution of single-particle reconstructions are discussed. We present reconstructions of six particles (DNA-dependent protein kinase catalytic subunit, alphaB-crystallin, the ribonucleoprotein vault, hepatitis A virus, adenovirus type 2, and the adenovirus type 12/alpha(v)beta5 integrin complex), which have a variety of symmetries (asymmetric to 60-fold) and a wide range of molecular masses (470 kDa to 150 MDa). In the case of icosahedral viruses, we have found that applying a "soft" mask to remove regions of disordered density improves the resolution given by the Fourier shell correlation 0.5 criterion. This masking procedure is also useful during refinement to improve the quality of the reference model and thus aid in precise alignment of the particle images. For asymmetric particles, we note that image classification, although often a necessary step to generate a first reconstruction, can limit the achievable resolution. The diameter of the particle and the available computational power can also affect the resolution, as can structural variability within the particle.

Adenoviridae↗

PubMed1999

High-resolution mapping of discrete representational areas in rat somatosensory cortex using blood volume-dependent functional MRI.

The present study documents the use of an iron oxide-based blood-pool contrast agent in functional magnetic resonance imaging to monitor activity-related changes in cerebral blood volume (CBV) resulting from peripheral sensory stimulation and the application of this technique to generate high-resolution functional maps. Rats, anesthetized with alpha-chloralose, were imaged during electrical stimulation (3 ms, 3 Hz, 3 V) of forelimb or hindlimb. Activation maps were generated by cross-correlation of the measured signal response and a square-wave function representative of the stimulus for each image pixel. Multislice imaging produced functional maps consistent with the known functional anatomy of rat primary somatosensory (S-I) cortex. Imaging with improved temporal resolution demonstrated rapid (<6 s) CBV increases which were sustained and relatively stable (coefficient of variation = 0.17 +/- 0.02) for forelimb stimulation periods of up to 5 min. Enabled by this sustained response we generated high-resolution (approximately 100 micrometer in-plane) functional maps showing discrete forelimb and hindlimb activation. This technique offers many advantages over other methods for the study of brain activity in the rat and has resolution sufficient to be useful in reorganization studies.

Animals↗

PubMed2002

Zoomed functional imaging in the human brain at 7 Tesla with simultaneous high spatial and high temporal resolution.

Functional neuroimaging in the human brain using noninvasive magnetic resonance methods has the potential of providing highly resolved maps of neuronal activation. Decreasing the voxel size and obtaining simultaneously high temporal resolution is a major challenge and is mainly limited by sensitivity. Here, signal-to-noise gains at high magnetic fields (7 Tesla) and an optimized surface coil setup are combined with a novel approach for zoomed functional imaging in the visual cortex. For echoplanar imaging, the acquisition time and segmentation was shortened fourfold by using a reduced field-of-view. An adiabatic outer-volume suppression method, BISTRO, was used to obliterate signal outside the area-of-interest achieving effective suppression even for inhomogeneous B1-fields. A single-shot acquisition was performed at submillimeter resolution in the human brain, while simultaneously maintaining a high temporal resolution of 125 ms. Functional studies with and without field-of-view reduction were performed. Activation and percent change maps were compared with respect to spatial extent, t values and percentage changes of the BOLD contrast. The detection of functional activation was found to be equal within the inter-series variability for the two acquisition schemes. Thus, single-trial BOLD responses were detected for the first time robustly at a 500 x 500 microm2 in plane and 250 ms temporal resolution, significantly expanding the possibilities of event-related functional imaging in the human brain. The magnetization transfer effect induced by the outer-volume suppression pulses was investigated and found to be increased during neuronal activity.

Algorithms↗

PubMed1999

Demonstration of differences in drug resistance by direct testing of DNA excision repair activity following standard and liposomal daunorubicin exposure in normal paediatric marrow using high resolution CLSM.

BACKGROUND: High resolution Confocal Laser Scanning Microscopy (CLSM) may be applied to testing of drug resistance in vitro in clinical setting. Rapid analysis of DNA damage by precise quantitation of excised DNA in bone marrow samples exposed to potential treatment moieties directly after isolation but the relative sensitivity of the integrated method is as yet untested. AIMS: To test the clinical applicability of SCGE/high resolution CLSM for differences in drug resistance in marrow cells. METHODS: Cells from normal bone marrow samples were exposed for identical periods and at 4 concentrations to either 1 hour of standard Daunorubicin (.5, 1, 1.5, 2 micrograms/ml) or 8 hours DaunoXome (courtesy of NeXstar Inc, USA) (.05, .1, .15, .2 microgram/ml). After 2 and 6 hours recovery, cells were harvested for SCGE, randomization, analysis of tail length, total excised DNA and fragment size distribution using high resolution CLSM. RESULTS: Tail length and fragment size distribution was not, but total excised DNA was significantly increased after 0.1 microgram/ml Liposomal Daunorubicin (DaunoXome) compared to 1.0 microgram/ml Daunorubicin. CONCLUSION: SCGE/high resolution CLSM effectively demonstrated differences in Daunorubicin resistance of human marrow cells to alternative formulations. The method has potential for use in clinical testing of neoplastic cell drug resistance.

Antibiotics, Antineoplastic↗

PubMed1994

Resolution of peritumoral brain edema following excision of meningioma.

We studied the resolution of peritumoral brain edema after meningioma excision. In twenty-nine patients with meningioma, the total volume of tumor and the peritumoral edema were measured planimetrically by serial CT scans and MRI with or without contrast enhancement. Four different patterns of postoperative resolution of hypodense volume on CT were observed: Group A: a large hypodensity rapidly decreased and disappeared, which may be related to the clearance of the real peritumoral edema in meningioma. Group B: a small hypodensity gradually disappeared. Group C: the hypodensity remained unchanged, which may result from the damaged brain tissue. Group D: the hypodensity progressively decreased but persisted, which may represent both the peritumoral edema and damaged brain tissue. We have calculated the resolution rate of edema fluid using the clearance curve of Groups A and D. The average resolution rate of edema fluid during the passage through 1 cm3 of the peritumoral white matter was 0.0493 ml/day. We speculate that 50% of edematous white matter, which presented as hypodensity on a CT scan, may be resolved in 4 days after total removal, and that 90% may be resolved in 14 days.

Blood-Brain Barrier↗

PubMed2001

High resolution FISH to delineate contiguous and small DNA sequences.

Somatic and meiotic metaphase, and pachytene chromosomes were subjected to DNA: DNA in situ hybridization to elucidate relative resolution of FISH signals for weak/contiguous hybridization sites. Hybridization with a '350 family' rye repetitive DNA probe pSc 200 characteristically differentiated the rye chromosome 5 from the rest of the complement on account of two small terminal homologous sites in the long arm, resolution of which is substantially improved using pachytene. Higher resolution of the two weak hybridization sites; a very small distal and a small proximal, is unequivocally demonstrated in the FISH painted 5RL examined at pachytene in the 5AS/5RL wheat background. Additionally this probe exhibits a large block of distal telomeric hybridization site in 5RS, followed by a more prominent proximal site homologous to '610 family' rye repetitive probe pSc 250. Precise denaturation - hybridization incubation and post hybridization stringency washing facilitates spatial resolution of contiguous repetitive rye probes pSc 200 and pSc 250, and physical localisation of small RFLP probe xpr 115 of wheat on barley chromosomes.

Chromosome Painting↗

PubMed1991

Characteristics of the normal central visual field measured with resolution perimetry.

The ring perimeter is an innovative device that measures the resolution threshold at 50 locations in the central visual field in approximately 6 min. This study was undertaken to define the characteristics of the resolution visual field in normal eyes using this instrument and to check the repeatability of the measurement. A total of 114 perimetrically naive, normal subjects aged from 20 to 79 years were prospectively enrolled; 55 randomly chosen subjects were tested twice, with a rest period of less than or equal to 60 s elapsing between examinations. The resolution threshold increased towards the periphery in all subjects. A small but significant decline in sensitivity occurred with age; this loss was greatest in the inner part of the field. Variability in threshold between subjects was not found to increase towards the periphery or in the upper field as compared with the lower. The field was highly repeatable when reexamined in the same subject. This study defines the resolution visual field in normal subjects, along with its inter- and intra-observer variability, and gives age-corrected predictive confidence intervals for each location of the field using the ring perimeter.

Adult↗

PubMed1994

High-resolution 1H NMR spectroscopy of aqueous humour from rabbits.

BACKGROUND: Detection of different substances in aqueous humour is important for evaluation of the disorders affecting the eye. The purpose of the present study was to apply high-resolution proton (1H) nuclear magnetic resonance (NMR) spectroscopy for extensive characterisation of the metabolites in the aqueous humour from rabbits. METHODS: High-resolution 1H NMR spectroscopy, including two-dimensional shift correlated (COSY) technique, was performed on aqueous humour from rabbits. RESULTS: More than 20 metabolites were simultaneously detected and identified in high-resolution 1H NMR spectra of aqueous humour from rabbits. Some of these were also quantified. CONCLUSION: High-resolution 1H NMR spectroscopy is a valuable method for simultaneous detection of many different metabolites in aqueous humour.

Animals↗

PubMed1993

Interpretation of high-resolution current source density profiles: a simulation of sublaminar contributions to the visual evoked potential.

Current source density (CSD) analysis provides an index of the location, direction, and density of transmembrane currents that arise with synchronous activation of neural tissue and that generate an evoked potential profile in the extracellular medium. In neocortex and other laminated structures, a simplified, one-dimensional CSD analysis can be computed by differentiation of voltages sampled at discrete points in a linear array. One-dimensional CSD analysis is a practical and accurate method for defining both regional activity patterns and neural generators of surface-recorded evoked and event-related potentials. In computing the CSD, common practices of differentiating across spatial grids of 200 microns or more and use of spatial smoothing routines help to reduce noise, but severely limit the spatial resolution available to the analysis. High-resolution CSD procedures (i.e., 3 point differentiation using a spatial grid of 100 microns or less) are more suited to identification of processes within individual cortical laminae or sublaminae, but can magnify the contributions of computational artifacts. Despite the inclusion of independent indices of cellular activity (e.g., multiunit activity), both high- and low-resolution analyses may indicate current source and sink configurations for which there is more than one plausible physiological interpretation. In the present study we examined the resolving capacity and pitfalls of common CSD procedures using simulated ensembles of current dipoles. These were positioned and oriented to model the depolarization of lamina 4C stellate cells and thalamocortical afferents in macaque striate cortex. Empirically, the surface N40 appears in association with a CSD configuration which includes current sinks within the thalamorecipient (stellate) subdivisions of lamina 4C and a large current source extending considerably below 4C. Dipole ensemble contributions to the CSD profile were computed and compared to physiological data from this region. Small asymmetries in activation of model stellate laminae were sufficient to produce substantial open field contributions. However, the best fit with empirical CSD profile was found when the simulation included contributions from thalamocortical axons, along with both open and closed field contributions from dual stellate cell sublaminae. High-resolution CSD profiles were shown to be interpretable when computational artifacts characteristic of closed and open fields were identified using a series of differentiation grids.

Afferent Pathways↗

PubMed1989

Measurement of resolution and recovery in recent generation positron tomographs.

In positron tomographic images, the ability to differentiate closely lying structures, the spillover of activity from a region into adjacent regions and the reduction in apparent isotope concentration in small structures are all dependent on spatial resolution. Resolution in the reconstructed image is affected by (i) detector size, (ii) the spatial sampling used (e.g. stationary, wobble), (iii) the amount of smoothing in the reconstruction process (or subsequent to reconstruction) and (iv) the image pixel size. Under ideal conditions, modern commercial tomographs can produce a reconstructed spatial resolution of 5 mm or less. However, this is rarely realizable in a clinical study due to the inadequacy of counting statistics and the amplification of statistical noise. In practice, a smoother filter has to be used. This paper presents a summary of practical measurements of spatial resolution, and the related count recovery, performed on recent generation positron tomographs. It is intended to contribute to the definition of methods of measuring these parameters which is part of an on going concerted action in positron tomography supported by the European Commission.

Image Processing, Computer-Assisted↗

PubMed1991

High-resolution magnetic resonance imaging of the interphalangeal joints of the hand.

High-resolution magnetic resonance imaging (MRI) of the interphalangeal joints of the fingers is being employed to study arthritis. To facilitate this research, a clear understanding of the structures visualisable by MRI is necessary. A gradient echo (GE) sequence was developed that produced good contrast between cartilage and other joint structures. These detailed images, with an in-plane resolution of 200 x 100 microns, enable resolution of three cartilage zones which can be interpreted as a superficial layer at the cartilage/cartilage interface, an intermediate layer and calcified cartilage in contact with bone; these correlate well with known anatomy. Further analysis of the images indicates that although a chemical shift artifact causes changes in the images at the field strength used (0.5 T), it does not cause enough distortion to necessitate suppression of the effect. Furthermore, the only detectable susceptibility artifact at these low field strengths was a loss of signal in bone trabeculae at the bone/cartilage interface. There is clearly potential in the study of the articular structures, in particular cartilage, in detail, using high-resolution MRI.

Adult↗

PubMed2004

Probe selection algorithm for oligonucleotide array-based medium-resolution genotyping.

Medium-resolution genotyping has the goal of distinguishing different subgroups instead of each element in a group. An oligonucleotide array provides an inexpensive, high-throughput method to identify differences in DNA sequence among individuals, which is fundamental for genotyping. As the cost and difficulty of designing and fabricating the oligonucleotide array dramatically increase with the number of probes used, it is therefore important to have a design with a minimum number of probes meeting the requirement of medium-resolution genotyping. The first algorithm for designing and selecting probes for oligonucleotide array-based medium-resolution typing is reported. The goal in deriving the algorithm was to select a minimum number of probes from a large probe set on the premise of minimum loss of resolution. The algorithm, which was based on entropy, conditional entropy and mutual information theory, was used to select the minimum number of probes from a large probe set. The algorithm was tested on a human leukocyte antigen (HLA) sequence data set Thirty probes were selected from 390 probes for HLA-A, and 60 probes were selected from 767 probes for HLA-B. Although the number of probes was reduced by almost ten times, the distinguishability was reduced only a little, by 0.45% (from 99.90% to 99.45%) for HLA-A and 0.27% (from 99.84% to 99.57%) for HLA-B, respectively. This is a satisfactory and practical result.

Algorithms↗

PubMed1994

Techniques and high resolution DNA size markers for pulsed field gel electrophoresis.

High resolution DNA size markers are described for pulsed field gel electrophoresis (PFGE). These markers provide resolution of 10-20 kbp over a size range from 10 kbp to more than 400 kbp and are produced by partial restriction digestion of lambda phage DNA concatemers (lambda ladder). High resolution markers extending to over 400 kbp are made by partial restriction digestion of lambda ladder embedded in agarose. Detailed methods are described for marker production and for DNA separation by contour-clamped homogeneous electric field (CHEF) electrophoresis. These markers and methods are useful for a variety of high resolution DNA mapping by PFGE.

Bacteriophage lambda↗

PubMed2001

Quality-control issues on high-resolution diagnostic monitors.

Previous literature indicates a need for more data collection in the area of quality control of high-resolution diagnostic monitors. Throughout acceptance testing, which began in June 2000, stability of monitor calibration was analyzed. Although image quality on all monitors was found to be acceptable upon initial acceptance testing using VeriLUM software by Image Smiths, Inc (Germantown, MD), it was determined to be unacceptable during the clinical phase of acceptance testing. High-resolution monitors were evaluated for quality assurance on a weekly basis from installation through acceptance testing and beyond. During clinical utilization determination (CUD), monitor calibration was identified as a problem and the manufacturer returned and recalibrated all workstations. From that time through final acceptance testing, high-resolution monitor calibration and monitor failure rate remained a problem. The monitor vendor then returned to the site to address these areas. Monitor defocus was still noticeable and calibration checks were increased to three times per week. White and black level drift on medium-resolution monitors had been attributed to raster size settings. Measurements of white and black level at several different size settings were taken to determine the effect of size on white and black level settings. Black level remained steady with size change. White level appeared to increase by 2.0 cd/m2 for every 0.1 inches decrease in horizontal raster size. This was determined not to be the cause of the observed brightness drift. Frequency of calibration/testing is an issue in a clinical environment. The increased frequency required at our site cannot be sustained. The medical physics division cannot provide dedicated personnel to conduct the quality-assurance testing on all monitors at this interval due to other physics commitments throughout the hospital. Monitor access is also an issue due to radiologists' need to read images. Some workstations are in use 7 AM to 11 PM daily. An appropriate monitor calibration frequency must be established during acceptance testing to ensure unacceptable drift is not masked by excessive calibration frequency. Standards for acceptable black level and white level drift also need to be determined. The monitor vendor and hospital staff agree that currently, very small printed text is an acceptable method of determining monitor blur, however, a better method of determining monitor blur is being pursued. Although monitors may show acceptable quality during initial acceptance testing, they need to show sustained quality during the clinical acceptance-testing phase. Defocus, black level, and white level are image quality concerns, which need to be evaluated during the clinical phase of acceptance testing. Image quality deficiencies can have a negative impact on patient care and raise serious medical-legal concerns. The attention to quality control required of the hospital staff needs to be realistic and not have a significant impact on radiology workflow.

Calibration↗

PubMed2004

Ultra-high-resolution X-ray structure of proteins.

The constant advances in synchrotron radiation sources and crystallogenesis methods and the impulse of structural genomics projects have brought biocrystallography to a context favorable to subatomic resolution protein and nucleic acid structures. Thus, as soon as such precision can be frequently obtained, the amount of information available in the precise electron density should also be easily and naturally exploited, similarly to the field of small molecule charge density studies. Indeed, the use of a nonspherical model for the atomic electron density in the refinement of subatomic resolution protein structures allows the experimental description of their electrostatic properties. Some methods we have developed and implemented in our multipolar refinement program MoPro for this purpose are presented. Examples of successful applications to several subatomic resolution protein structures, including the 0.66 angstrom resolution human aldose reductase, are described.

Aldehyde Reductase↗