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PubMed1990

Banding resolution of amniotic cell chromosome preparations for prenatal diagnosis.

Fifty consecutive routine female karyotypes of amniotic fluid cells were analyzed to determine: (1) the banding resolution of each metaphase in routine preparations, (2) whether all haploids in the same metaphase had the same level of banding resolution, and (3) which chromosomes showed the least/most variation in regard to banding resolution. For each karyotype, the number of dark bands for each right-hand-side chromosome were counted and compared with an ideogram (ISCN 1985). Banding resolution was separated into three groups: 400-band, 550-band, and 850-band. It was found that all 50 metaphases showed at least a 400-band resolution, and 1 metaphase showed a banding resolution between a 550-band and 850-band level. Although the chromosome preparations were sufficient for routine purposes, additional effort and banding techniques are required to detect small chromosomal abnormalities that require a higher level of resolution.

Amniotic Fluid↗

PubMed1986

Effects of centre-of-rotation shift on contrast and spatial resolution of the SPECT system.

The effects of centre-of-rotation (COR) shift of the rotating camera-based single photon emission commuted tomography (SPECT) system on tomographic images were investigated quantitatively. Contrast and spatial resolution variations as a function of COR shift were measured for all shifts less than 1 pixel, in steps of 0.1 pixels. This region encompasses the typical COR variations. The behaviour of these indices was analysed. In addition, the validity of the limit imposed on the allowable COR shift, as utilized in the current SPECT quality control protocols, was examined. The value of either +/- 0.5 or +/- 0.25 pixels is presently in use as the limit. From this study, both limits result in the loss of resolution. The resolution loss will increase the uncertainties of volume and concentration measurements. For a COR shift of 0.25 pixels, the spatial resolution loss is approximately 5%. This small spatial resolution loss may not affect the qualitative evaluation of tomographic images. However, any loss of resolution should be avoided. For a COR shift of 0.1 pixels, there is no observed loss of resolution. This value should be a desirable choice as the maximum allowable COR shift since it ensures the optimum performance of the SPECT system.

Rotation↗

PubMed1988

Visual resolution of macaque retinal ganglion cells.

1. The visual resolving ability of different types of macaque retinal ganglion cells was estimated at different retinal eccentricities, by measuring the amplitude of modulated responses to black-white gratings of spatial frequencies near the resolution limit for each cell. 2. The resolving ability of tonic, spectrally opponent ganglion cells was usually similar to that of phasic, non-opponent ganglion cells at similar eccentricities, except that at eccentricities greater than 10 deg some tonic ganglion cells with remarkably high resolution (up to ca. 15 cycles/deg) were found. Our cell sample was limited within the central 2 deg of the visual field, however. 3. Only a small proportion of phasic ganglion cells showed an increase of mean firing level to gratings near the resolution limit. The maintained firing of tonic ganglion cells was higher than that of phasic ganglion cells. 4. With red-black or green-black gratings, the resolution of phasic ganglion cells was unaffected. For red or green on-centre ganglion cells, a marked deterioration of resolving ability occurred when the grating was of a colour to which a cell responded poorly (green-black gratings for red on-centre cells, and red-black gratings for green on-centre cells). A slight improvement in resolving ability occurred when the grating was of an excitatory colour. 5. For a sub-sample of cells, we compared resolution limit with centre size as determined from area-threshold curves. For both phasic and tonic ganglion cells, resolution limit (the period length just resolved) was about half the centre diameter, as is the case for cat ganglion cells. This implies that the centre sizes of phasic and tonic monkey ganglion cells are similar at most eccentricities. 6. We attempt to relate these results to primate retinal anatomy and visual resolution, determined behaviourally.

Action Potentials↗

PubMed1992

Intensity and frequency resolution: masking of absolute identification and fixed and roving discrimination.

Auditory intensity and frequency resolution were studied in three paradigms under masking conditions. Absolute identifications of single stimuli (one-interval paradigm) and 2IFC judgments of fixed- and roving-level pairs of stimuli (two-interval paradigm) were obtained from the same experienced observers. Judgments were made under optimal (no mask) conditions, in the presence of a broadband noise mask (simultaneous mask), and when the stimulus(i) to be judged were either preceded (forward mask) or followed (backward mask) by a broadband noise mask. Substantial masking of intensity resolution was found in all mask conditions. Only a simultaneous mask affected frequency resolution. In the no mask condition, performance was best for fixed-level (or frequency) 2IFC discrimination, followed by roving-level (frequency) 2IFC, and finally absolute identification. These differences were maintained under masking for frequency resolution, but not for intensity resolution. The results are discussed in terms of the Braida and Durlach (1988) model of intensity resolution. A similar model is suggested for frequency resolution with differences suggested by the differences in neural coding of sound intensity and frequency.

Auditory Perception↗

PubMed1987

Efficient resolution of replicated poxvirus telomeres to native hairpin structures requires two inverted symmetrical copies of a core target DNA sequence.

The terminal hairpin sequences of the linear double-stranded DNA genome of the leporipoxvirus Shope fibroma virus (SFV) has been cloned in Saccharomyces cerevisiae and in recombination-deficient Escherichia coli as a palindromic insert within circular plasmid vectors. This sequence configuration is equivalent to the inverted repeat structure detected as a telomeric replicative intermediate during poxvirus replication in vivo. Previously, it has been shown that when circular plasmids containing this palindromic insert were transfected into SFV-infected cells, efficient replication and resolution generated linear minichromosomes with bona fide viral hairpin termini (A. M. DeLange, M. Reddy, D. Scraba, C. Upton, and G. McFadden, J. Virol. 59:249-259, 1986). To localize the minimal target DNA sequence required for efficient resolution, a series of staggered unidirectional deletions were constructed at both ends of the inverted repeat. Analyses of the resolution efficiencies of the various clones indicate that up to 240 base pairs (bp) centered at the symmetry axis were required for maximal resolution to minichromosomes. To investigate the role of the AT-rich central axis sequences, which in SFV include 8 nonpalindromic bp, a unique AflII site at the symmetry axis was exploited. Bidirectional deletions extending from this AflII site and insertions of synthetic oligonucleotides into one of the deletion derivatives were constructed and tested in vivo. The efficiency with which these plasmids resolved to linear minichromosomes with hairpin termini has enabled us to define the minimal target DNA sequence as two inverted copies of an identical DNA sequence between 58 and 76 bp in length. The nonpalindromic nucleotides, which, after resolution, constitute the extrahelical residues characteristic of native poxviral telomeres, were not required for resolution. The close resemblance of the SFV core target sequence to the analogous region from the orthopoxvirus vaccinia virus is consistent with a conserved mechanism for poxviral telomere resolution.

DNA↗

PubMed1990

Frequency selectivity and temporal resolution in patients with various inner ear disorders.

Both frequency selectivity and temporal resolution were examined in patients with various inner ear disorders. These included noise-induced hearing loss (n = 24), Menière's disease (n = 16), sudden deafness (n = 25), toxic inner ear damage (n = 14), presbyacusis (n = 38) and degenerative progressive inner ear hearing loss (n = 8). To facilitate quantitative comparison, various factors were introduced, namely, frequency resolution factor (FRF), temporal resolution factor (TRF) and a combined resolution factor (FTRF). The FRF of normal-hearing subjects in background noise conditions was found to be approximately 20% less than in comparative test conditions without noise, whereas the TRF of normal-hearing persons tested under background noise conditions showed a remarkable increase (factor 3). The frequency resolution performance and/or the temporal resolution performance were found to be impaired in all patient groups with inner ear hearing loss. This is particularly noticeable for temporal resolution in test conditions involving the addition of background noise. It can be concluded that in such cases, speech discrimination can be seriously jeopardized.

Hearing↗

PubMed1989

Chromosome preparation and high resolution banding techniques. A review.

High resolution banding techniques enable detection of chromosome rearrangements even within major bands. Banded chromosomes prepared for light microscopic studies of intact metaphase plates are, however, highly modified structures compared with native chromosomes, and the high resolution banding techniques only seem possible because the following methods were standardized and combined. The use of colcemid, which prevents formation of the spindle and thereby collects cells at the metaphase-anaphase border, is routinely used for chromosome preparations. For high resolution banding studies, short exposure time and concentrations near the threshold value have been recommended by several authors. Several agents interfere with chromosome contraction processes, but only a few have had a lasting influence on high resolution banding studies. The most used agents are ethidium bromide, actinomycin D, and Hoechst 33258, which all partially inhibit chromosome contraction. Treatment with hypotonic solutions induces swelling of animal cells, and the methanol in the fixative denatures and precipitates protein by dehydration. The acetic acid coagulates nucleoproteins and causes swelling of the cells. The fixative penetrates the cells rapidly and preserves the chromosome structure. To obtain long segmented chromosomes suitable for high resolution banding hypotonic treatment with .075 M KCl, frequent changes of fixative and overnight fixation at 4 degrees C have been recommended. The use of cell synchronization, 5-bromodeoxyuridine incorporation into DNA, and fluorochrome-photolysis Giemsa (FPG)-staining have improved the quality of high resolution banding. Synchronization techniques, which select for lymphocyte populations in early divisions, provide excellent materials for chromosome preparations and induction of high resolution banding. The banding techniques seem to enhance differences already present in the chromosomes, and the differential Giemsa staining has recently been explained by interactions between the hydrophobic dye complex, the supercoiled DNA helix, and the denaturated histone core of the nucleosomes.

Animals↗

PubMed1975

Effect of positron range on spatial resolution.

The effect of beta+ range on spatial resolution of imaging systems employing the detection of 511-keV annihilation radiation was determined by measuring the variation in the line-spread functions (LSFs) of positron-emitting radionuclides of 64Cu, 11C, and 15O as compared with the 514-keV gamma-ray emitter 85Sr. These radionuclides have maximum beta+ energies of 0.656, 0.960, and 1.72 MeV, respectively. The LSFs were measured in a tissue-equivalent phantom with high-resolution (approximately 2.4 mm FWHM) and low-resolution (approximately 8.8 mm FWHM) straightbore collimators coupled to a NaI(Tl) detector. Theoretical LSFs for the beta+ ranges were also calculated and convolved with the 85Sr LSF to yield the predicted LSFs for 11C and 15O. The high-resolution study showed a 0% and 2.3% increase in the full-width half-maximum (FWHM) and full-width tenth-maximum (FWO.1M) for the low-energy beta+ of 64Cu and a 37% (FWHM) and 52% (FWO.1M) increase for the high energy beta+ of 15O as compared with 85Sr. However, when the system resolution was decreased to 8.8 mm FWHM, the 64Cu showed no change at FWHM or FWO.1M and the 15O showed a 2.3% (FWHM) and 7.8% (FWO.1M) relative to 85Sr. The predicted LSFs were in good agreement with the experimental. These data indicate that the effect of beta+ range on spatial resolution is minimal unless the beta+ energy is larger than or equal to 1.5 MeV and the system resolution is on the order of a few millimeters.

Carbon Radioisotopes↗

PubMed1992

Phosphorylation-induced conformational changes in the phosphorylase ab hybrid as revealed by resolution of pyridoxal 5'-phosphate with imidazole citrate and cysteine.

The accessibility of pyridoxal 5'-phosphates of the phosphorylase ab hybrid to resolution by imidazole citrate and cysteine was studied and compared with that of the b and a forms. Promotion of resolution of phosphorylated forms by raising the temperature or in the presence of glycogen indicates that the resistance of phosphorylase a and ab to resolution at 0 degrees C is due rather to their tetrameric state than their phosphorylation-related active conformation. The pattern of resolution of the ab hybrid was similar to that of the a and differed from that of the b forms in that it occurred at 30 degrees C and 37 degrees C but not at 0 degrees C, moreover, it did not show first-order kinetics. On the other hand, inhibition of resolution by ligands binding to the nucleotide site of phosphorylase reflected an intermediate sensitivity of the ab form between that of the b and a forms. We conclude that partial phosphorylation of phosphorylase b elicits conformational change(s) in both subunits which influence the monomer-monomer interactions and resolution of pyridoxal 5'-phosphates. Resistance of ab hybrid to monomerizing agents as imidazole citrate, comparable to that of other forms, argues for its stability, ruling out its reshuffling into mixtures of phosphorylase b and a.

Adenosine Monophosphate↗

PubMed1987

Resolution of the enantiomers of various alpha-substituted ornithine and lysine analogs by high-performance liquid chromatography with chiral eluant and by gas chromatography on Chirasil-Val.

A reversed-phase high-performance liquid chromatography method, with L-proline and copper as chiral mobile phase, is described for the enantiomeric resolution of various alpha-substituted ornithine and lysine analogs. Although ornithine gives no separation with the chiral eluant used, excellent resolutions are obtained for various alpha-alkyl-, alpha-halogenomethyl-, alpha-vinyl-, and alpha-ethynyl-substituted ornithines. Similar separations are also observed for the dehydroornithine and lysine analogs. Gas chromatography on a chiral stationary phase, Chirasil-Val, allows the resolution of the ornithine and lysine analogs after derivatization into the monofluoroacyl derivatives of their corresponding lactams. No resolution or only a poor resolution is obtained by GC on Chirasil-Val for the dehydroornithine analogs as their di-N-perfluoroacyl alkyl esters. The chiral eluant HPLC procedure is easily scaled up for the semipreparative resolution of several ornithine analogs, i.e., alpha-fluoromethylornithine, alpha-difluoromethylornithine, alpha-chlorofluoromethylornithine, and alpha-fluoromethyldehydroornithine, which are known as potent ornithine decarboxylase inhibitors in vitro and in vivo.

Chromatography, Gas↗

PubMed1987

High resolution autoradiography at the regional topographic level with [14C]2-deoxyglucose and [3H]2-deoxyglucose.

After injection of 2-deoxyglucose (2-DG) labeled with tritium or carbon-14, autoradiograms were produced by thaw-mounting 4 micron frozen sections of rat brains on nuclear emulsion-coated slides. The results show that the distribution of radioactivity among different brain regions was similar and that the resolution at the regional topographical level was virtually identical for both compounds. The resolution obtained with the thaw-mounting of thin frozen sections onto nuclear emulsion was considerably greater than the resolution demonstrated in published results in the literature, when carbon-14 or tritium-labeled 2-deoxyglucose were used with 20 micron frozen sections and X-ray film or tritium-sensitive film. The results indicate that section thickness, detection medium and intimacy of contact between section and photographic emulsion influence resolution. At the regional level, the detection medium apparently influences resolution to a greater extent than the energy differences of the beta particles emitted from 14C or 3H. Although diffusion of radiolabeled 2-deoxyglucose and metabolites during the thaw-mounting process precludes single cell resolution of these autoradiograms, the improvement of visualizing regional topographic detail demonstrates that the described technique is a valuable approach with which to study regional 2-DG uptake.

Animals↗

PubMed1992

Emission microscopy and related techniques: resolution in photoelectron microscopy, low energy electron microscopy and mirror electron microscopy.

A unified treatment of the resolution of three closely related techniques is presented: emission electron microscopy (particularly photoelectron microscopy, PEM), low energy electron microscopy (LEEM), and mirror electron microscopy (MEM). The resolution calculation is based on the intensity distribution in the image plane for an object of finite size rather than for a point source. The calculations take into account the spherical and chromatic aberrations of the accelerating field and of the objective lens. Intensity distributions for a range of energies in the electron beam are obtained by adding the single-energy distributions weighted according to the energy distribution function. The diffraction error is taken into account separately. A working resolution is calculated that includes the practical requirement for a finite exposure time, and hence a finite non-zero current in the image. The expressions for the aberration coefficients are the same in PEM and LEEM. The calculated aberrations in MEM are somewhat smaller than for PEM and LEEM. The resolution of PEM is calculated to be about 50 A, assuming conventional UV excitation sources, which provide current densities at the specimen of 5 x 10(-5) A/cm2 and emission energies ranging up to 0.5 eV. A resolution of about 70 A has been demonstrated experimentally. The emission current density at the specimen is higher in LEEM and MEM because an electron gun is used in place of a UV source. For a current density of 5 x 10(-4) A/cm2 and the same electron optical parameters as for PEM, the resolution is calculated to be 27 A for LEEM and 21 A for MEM.

Mathematics↗

PubMed1988

Silicone intraocular lens resolution in air and in water.

The resolution efficiencies of 31 biconvex silicone intraocular lenses, ranging in power from 16.0 to 23.5 diopters, were tested in air and in water to see if a predictable relationship existed as previously reported with polymethylmethacrylate lenses. Resolution efficiency is defined as the percentage ratio of the actual resolving power of a lens to that of a perfect lens of the same focal length which is only limited in resolution by diffraction. The lenses ranged from 29% to 58% resolution efficiency in air. No lenses exhibiting multiple images were included. All 31 lenses achieved at least 73% resolution efficiency in water, and one lens achieved 82%. Based on these findings, a biconvex silicone lens that exceeds 30% resolution efficiency in air and does not produce multiple images can perform near its diffraction limit when implanted in the eye.

Air↗

PubMed1987

Video-endoscopes in comparison with fiberscopes: quantitative measurement of optical resolution.

The subject of this study was the optical performance of video-endoscopic systems (VE) in terms of maximal resolving power and resolvable picture elements. Olympus, Toshiba/Machida, Fujinon, and Welch Allyn video gastroscopes were tested. A GIF Q 10 fiberscope from Olympus was also included for comparison. The resolution measurements were made at various distances using two independent methods--electronic analysis of the TV signal, and visual evaluation of the resolution, of a standardized test target. The results obtained with the two methods were in perfect agreement. The resolution of fine details clearly depends on the distance between the distal end and the target because of decreasing image scale. Depending on the individual optical design, the various VE's show maxima at different distances. At shorter distances, the image is degraded by defocusing. An optimal distance which is as small as possible is desired for clinical routine. Apart from the fiberscope this requirement is best met by the Fuji system. The greatest resolution is obtained with the Toshiba system but at the cost of the viewing angle which is the smallest of all the systems. Fuji combines relatively high resolution with a large viewing angle. Because of the widely varying viewing angle a comparison based solely on resolution cannot represent the true imaging capability of the system. We therefore eliminated purely optical parameters and calculated the number of resolvable picture elements per line. We regard this number to be a fair characterization of both TV and fiber systems.

Fiber Optic Technology↗

PubMed1987

Influence of zoom factor on centre-of-rotation of the SPECT system and on the resolution of tomographic images.

The influence of zooming on centre-of-rotation (COR) of the rotating camera-based SPECT system and on the resolution of tomographic images was investigated. COR values were measured as a function of zoom factor varying from x1.0 to x3.0. Measurements were performed using two SPECT systems collimated with three collimators. In this study, the COR was observed to shift linearly with zoom factor. The influence of zooming on COR depends on the location and the deviation of the COR from the axis-of-rotation (AOR). We recommend that the deviation of the COR from the AOR be kept to within 0.4 pixels corresponding to 2.4 mm. Unlike zooming, the magnitude of COR shift is uncorrelated to the type of collimator used. Contrast and spatial resolution variations of the tomographic images were quantitated as a function of COR shift for clinical data acquired using no zoom factor and a zoom factor of 2.50. The resolution indices were observed to be more sensitive to COR shift for clinical data acquired using no zoom factor. The sensitivity appeared to be inversely dependent on the zoom factor utilized. However, zooming was found not to preserve the same resolution degradation. Optimum resolution images can be reconstructed by requiring the COR be determined and clinical study be acquired using the same zoom factor and collimator. For qualitative evaluation of tomographic images, the zoom factor requirement can be removed provided that the deviation of the COR from the AOR is within the recommended limit. This limit allows for a maximum spatial resolution degradation of 5%.(ABSTRACT TRUNCATED AT 250 WORDS)

Image Enhancement↗

PubMed1992

High-resolution scanning electron microscopy of frozen-hydrated cells.

Cryo-fixed yeast Paramecia and sea urchin embryos were investigated with an in-lens type field-emission SEM using a cold stage. The goal was to further develop and investigate the processing of frozen samples for the low-temperature scanning electron microscope (LTSEM). Uncoated frozen-hydrated samples were imaged with the low-voltage backscattered electron signal (BSE). Resolution and contrast were sufficient to visualize cross-fractured membranes, nuclear pores and small vesicles in the cytoplasm. It is assumed that the resolution of this approach is limited by the extraction depth of the BSE which depends upon the accelerating voltage of the primary beam (V0). In this study, the lowest possible V0 was 2.6 kV because below this value the sensitivity of the BSE detector is insufficient. It is concluded that the resolution of the uncoated specimen could be improved if equipment were available for high-resolution BSE imaging at 0.5-2 kV. Higher resolution was obtained with platinum cryo-coated samples, on which intramembranous particles were easily imaged. These images even show the ring-like appearance of the hexagonally arranged intramembranous particles known from high-resolution replica studies. On fully hydrated samples at high magnification, the observation time for a particular area is limited by mass loss caused by electron irradiation. Other potential sources of artefacts are the deposition of water vapour contamination and shrinkage caused by the sublimation of ice. Imaging of partially dehydrated (partially freeze-dried) samples, e.g. high-pressure frozen Paramecium and sea urchin embryos, will probably become the main application in cell biology. In spite of possible shrinkage problems, this approach has a number of advantages compared with any other electron microscopy preparation method: no chemical fixation is necessary, eliminating this source of artefacts; due to partial removal of the water additional structures in the cytoplasm can be investigated; and finally, the mass loss due to electron beam irradiation is greatly reduced compared to fully frozen-hydrated specimens.

Animals↗

PubMed1989

High resolution scanning electron microscopy of the cell.

The scanning electron microscope (SEM) has become a powerful tool for ultrastructural research with improvement of the instrument's resolution and progress in specimen preparation techniques. With regard to resolution, it has been improved step-by-step in this decade and, in 1985, an ultra-high resolution SEM (UHS-T1) was developed, with a resolution of 0.5 nm. Concerning specimen preparation, the osmium-DMSO-osmium method, which is effective for revealing intracellular structures, has come to be widely used. Techniques for observing smaller objects, such as bacteriophages, viruses, and biological macromolecules, have also been devised in recent years. As a result of these preparation techniques and the availability of the ultra-high resolution SEM, the application of SEM in biology is expanding rapidly. In this paper, an outline of the ultra-high resolution SEM, techniques for specimen preparation, findings of some biological materials by these techniques, and guidelines to making the specimens, are described.

Animals↗

PubMed1967

Visual resolution when light enters the eye through different parts of the pupil.

1. Threshold contrasts for resolution of sinusoidal gratings imaged on to the retina through a decentred 2 mm pupil were measured.2. No loss in resolution was found when the pupil was decentred parallel to the lines of the gratings. A loss in resolution by a factor of 3 occurred when the pupil was decentred by 3 mm perpendicular to the lines of the gratings.3. The effects of focus on the threshold contrast for a grating viewed through a centred and decentred pupil were used to show that at least a portion of the loss in resolution is due to optical aberrations.4. Using a neon-helium gas laser as a coherent light source, interference fringes were produced on the retina directly. Threshold contrasts for resolution of the fringes were determined for different positions of entry of the beams of light through the pupil. When the Stiles-Crawford brightness effect was compensated for, no loss in resolution was found to occur for decentred entry of the beams.5. It is concluded that the off-axis loss of visual acuity is wholly due to optical aberrations in the eye.6. The ratios between the threshold contrasts for sinusoidal gratings and for interference fringes are used to calculate the optical transfer functions of the off-axis aberrations of the eye.

Humans↗