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Quantitation of cell area on glass and fibronectin-coated surfaces by digital image analysis.

By using digital image processing and analysis, two procedures were developed to rapidly measure the projected area of a field of adherent 3T3 fibroblasts without staining of cell borders. The cell area of newly attached and rounded cells with well-resolved borders was obtained by a gray value thresholding procedure. For cells that had undergone an appreciable degree of spreading, cell boundaries were less distinct and a nonlinear spatial Sobel filter was used, followed by thresholding. For both procedures, linear relations were observed between cell areas obtained from image analysis and cell areas obtained by tracing. The areas of a population of traced cells were not statistically different from the area distribution obtained by using the standard curves for the processed images. Uncertainty in the estimated mean area depended only upon the number of cells examined. Approximate numbers of cells required to obtain estimates of the mean are calculated. As an application of these procedures, cell areas were measured for 3T3 cells attached to glass and fibronectin-coated surfaces and were found to be significantly larger for cells spreading on fibronectin-coated glass than on glass alone. Increased cell area during spreading on fibronectin-coated surfaces was proportional to increased cell adhesivity after exposure to a shear stress of 58 dyn/cm2.

Cell Adhesion↗

Microdistribution of substratum-bound ligands affects cell function: hepatocyte spreading on PEO-tethered galactose.

Cell functions are regulated by signal transduction processes triggered by binding interactions with extracellular ligands, including those coupled to insoluble matrices as well as those diffusing in free solution. Whereas receptor interactions with freely diffusible soluble ligands are quantitatively governed by the mean ligand concentration, interactions with spatially constrained substratum-bound ligands may be affected not only by the mean ligand concentration, but also by the ligand spatial microdistribution. To probe this hypothesis we have generated surfaces presenting galactose ligands at different concentrations and different capabilities for micromobility--and thus spatial microdistribution--by means of polyethylene oxide tethering and assayed for hepatocyte spreading mediated by asialoglycoprotein receptor/cytoskeleton linkages. We demonstrate that spreading is not uniquely determined by the mean galactose concentration presented by the culture substrate. Rather, the ability of primary hepatocytes to spread is additionally specified by a combination of ligand concentration and tether length. Our results suggest spreading results when monovalent ligands possess sufficient mobility to form effectively multivalent bonds with the receptor; that is, when the substratum-bound ligands to cluster in spatial microdomains.

Animals↗

Identification of the cholera diffusion process in Ibadan, 1971.

The paper tries to examine and identify which spatial diffusion process was responsible for generating the pattern of cholera diffusion (an epidemic spread which was apparently wave-like) within Ibadan City in 1971. In this paper one of Moran's statistics, the BW join-count measure of spatial autocorrelation is employed. Five different plannar graphs are used in the study. The results show that contagion was apparent on the various models of spatial processes employed and on their different combinations. But it is the radial contact diffusion which was discovered to be most important in the spread of the epidemic. But even, though such a radial contact diffusion process was discovered to be the most important, during the advance and peak phases of the epidemic wave; as the epidemic intensity rose to a spread phase, a mixture of the various models became a best contributor to the contagion.

Cholera↗

Virus spread and initial pathological changes in the nervous system in genital herpes simplex virus type 2 infection in mice. A correlative immunohistochemical, light and electron microscopic study.

Mice were infected by the vaginal route with the MS strain of herpes simplex virus type 2 (HSV-2). Serial vaginal cultures were used to confirm infection and to select mice for this study. Two mice were killed by perfusion on days 2-6 post infection (p.i.) and lumbar and sacral cord with cauda were fixed and embedded for electron microscopy. Semithin Epon-sections were stained for viral antigen using a rabbit anti-HSV-2 antiserum and the Avidin-Biotin (ABC) method. Thin sections from antigen-positive blocks were examined by electron microscopy, and the number and types of infected cells detected by these two methods were compared. A good correlation was found between detection of infected cells by these methods. Infected cells included neurons of dorsal root ganglia and spinal cord, satellite cells of dorsal root ganglia, non-myelinating Schwann cells, astrocytes, oligodendrocytes and arachnoidal cells. Infected cells were first detected in the cauda on day 3 p.i. and in the spinal cord on day 5 p.i. The temporal and spatial distribution of infected cells was consistent with neural spread to and within the CNS. The pathological lesions showed a good correlation with the distribution and number of infected cells and are probably due to a direct virus effect. The similar sensitivity of the Epon-ABC method to electron microscopy in detecting infected cells indicates that this method may have useful applications in both experimental and diagnostic work.

Animals↗

Nonlinear charge spreading visualized in voltage-controlled lateral superlattices.

Voltage-controlled lateral potential superlattices are used to dramatically increase the lifetime of photogenerated carriers in a quantum well. These long lifetimes, together with the ability to deliberately trigger radiative recombination, enable us to directly visualize the spreading character of nonlinear Maxwell relaxation of 2D charges along narrow channels. Our system allows for temporal and spatial resolution of Maxwell kinetics, usually a very fast process and difficult to observe. The observed spreading dynamics of a 2D hole plasma is in perfect agreement with our nonlinear model.

Journal Article↗

Development of high quantum efficiency flat panel detectors for portal imaging: intrinsic spatial resolution.

Recently developed flat panel detectors have been proven to have a much better image quality than conventional electronic portal imaging devices (EPIDs). They are, however, not yet the ideal systems for portal imaging application due to the low x-ray absorption, i.e., low quantum efficiency (QE), which is typically on the order of 2-4% as compared to the theoretical limit of 100%. The QE of current flat panel systems can be improved by significantly increasing the thickness of the energy conversion layer (i.e., amorphous selenium or phosphor screen). This, however, will be at the expense of a decrease in spatial resolution mainly due to x-ray scatter in the conversion layer (and also the spread of optical photons in the case of phosphor screen). In this paper, we investigate theoretically the intrinsic spatial resolution of a high QE flat panel detector with a new energy conversion layer that is much denser and thicker than that of current flat panel systems. The modulation transfer function (MTF) of the system is calculated based on a theoretical model using a novel approach, which uses an analytical expression for absorbed dose. It is found that if appropriate materials are used for the conversion layer, then the intrinsic MTF of the high QE flat panel is better than that of current EPIDs, and in addition they have a high QE (e.g., approximately 60%). Some general rules for the design of the conversion layer to achieve both high QE and high resolution as well as high DQE are also discussed.

Computer Simulation↗

Dynamics of keratin filaments and the intermediate filament distribution center during shape change in PtK1 cells.

Reorganization of intermediate filaments during cell spreading is examined by immunofluorescence, electron microscopy, and time-lapse video microscopy. A juxtanuclear cap, believed to correspond to the intermediate filament distribution center, was observed to be spatially related to the organization of the intermediate filament network as cells spread. A keratin cap was observed, which appeared spontaneously in motile PtK1 cells. Cap formation may be a consequence of retraction of intermediate filaments from the cytoplasm as cells move. The position of this juxtanuclear cap is related to the direction of movement, located on the side of the nucleus near the advancing edge of the cell. As the cell spreads, the cap disappears as the keratin filament network returns to the cytoplasm. Evidence presented here is consistent with the hypothesis that the distribution center mediates keratin filament organization during cell shape change.

Animals↗

An edge spread technique for measurement of the scatter-to-primary ratio in mammography.

An experimental measurement technique that directly measures the magnitude and spatial distribution of scatter in relation to primary radiation is presented in this work. The technique involves the acquisition of magnified edge spread function (ESF) images with and without scattering material present. The ESFs are normalized and subtracted to yield scatter-to-primary ratios (SPRs), along with the spatial distributions of scatter and primary radiation. Mammography is used as the modality to demonstrate the ESF method, which is applicable to all radiographic environments. Sets of three images were acquired with a modified clinical mammography system employing a flat panel detector for 2, 4, 6, and 8 cm thick breast tissue equivalent material phantoms composed of 0%, 43%, and 100% glandular tissue at four different kV settings. Beam stop measurements of scatter were used to validate the ESF methodology. There was good agreement of the mean SPRs between the beam stop and ESF methods. There was good precision in the ESF-determined SPRs with a coefficient of variation on the order of 5%. SPRs ranged from 0.2 to 2.0 and were effectively independent of energy for clinically realistic kVps. The measured SPRs for 2, 4, and 6 cm 0% glandular phantoms imaged at 28 kV were 0.21+/-0.01, 0.39+/-0.01, and 0.57+/-0.02, respectively. The measured SPRs for 2, 4, and 6 cm 43% glandular phantoms imaged at 28 kV were 0.20+/-0.01, 0.35+/-0.02, and 0.53+/-0.02, respectively. The measured SPRs for 2, 4, and 6 cm 100% glandular phantoms imaged at 28 kV were 0.22+/-0.02, 0.42+/-0.03, and 0.88+/-0.08, respectively.

Biophysical Phenomena↗

The number and spatial distribution of IP3 receptors underlying calcium puffs in Xenopus oocytes.

Calcium puffs are local Ca(2+) release events that arise from a cluster of inositol 1,4,5-trisphosphate receptor channels (IP(3)Rs) and serve as a basic "building block" from which global Ca(2+) waves are generated. Important questions remain as to the number of IP(3)Rs that open during a puff, their spatial distribution within a cluster, and how much Ca(2+) current flows through each channel. The recent discovery of "trigger" events-small Ca(2+) signals that immediately precede puffs and are interpreted to arise through opening of single IP(3)R channels-now provides a useful yardstick by which to calibrate the Ca(2+) flux underlying puffs. Here, we describe a deterministic numerical model to simulate puffs and trigger events. Based on confocal linescan imaging in Xenopus oocytes, we simulated Ca(2+) release in two sequential stages; representing the trigger by the opening of a single IP(3)R in the center of a cluster for 12 ms, followed by the concerted opening of some number of IP(3)Rs for 19 ms, representing the rising phase of the puff. The diffusion of Ca(2+) and Ca(2+)-bound indicator dye were modeled in a three-dimensional cytosolic volume in the presence of immobile and mobile Ca(2+) buffers, and were used to predict the observed fluorescence signal after blurring by the microscope point-spread function. Optimal correspondence with experimental measurements of puff spatial width and puff/trigger amplitude ratio was obtained assuming that puffs arise from the synchronous opening of 25-35 IP(3)Rs, each carrying a Ca(2+) current of approximately 0.4 pA, with the channels distributed through a cluster 300-800 nm in diameter.

Animals↗

1H NMR spectroscopic imaging of the monkey brain using binomial water suppression in a stimulated-echo sequence.

A new proton, two-dimensional pulse sequence for 1H NMR spectroscopic imaging (chemical shift imaging) was tested in phantoms and in the monkey brain. The pulse sequence consisted of one binomial chemically selective pulse and two spatially selective pulses in the stimulated-echo sequence. The point-spread function (which is influenced by k-space filtering and the number of phase-encoded steps) of a 1 mm source phantom was measured using a 16 x 16 spatial matrix and was found to have a FWHM of 10 mm (100 mm field of view) with very little rippling outside the main lobe. The binomial excitation profile was measured in order to correct the NMR intensity for the variable flip angles. Spectroscopic images were measured in the monkey brain with a 15 mm slice thickness and a 16 x 16 spatial matrix. Proton spectra derived from the brain contained sharp resonances of choline, creatine and N-acetyl aspartate with minimal lipid contamination. Proton spectra derived from the subcutaneous fat and adipose tissue behind the eyes contained large lipid resonances.

Animals↗

Use of retroillumination to visualize optical aberrations caused by tear film break-up.

PURPOSE: The aim of the current study was to develop quantitative methods to assess optical aberrations caused by tear film disruption. METHODS: We used standard fluorescein imaging (FL) and a novel retroillumination (RI) method to image tear film disruption in 12 eyes. Using a clinical slit lamp biomicroscope, we alternated between widefield blue and narrow-beam white light to obtain an interleaved series of FL and RI images of the time course and pattern of tear film break-up. We developed an optical analysis that indicates that the RI image should be proportional to the spatial derivative of the FL image. Intensity fluctuations in the RI images are due to thickness changes in the tear film, whereas intensity fluctuations in FL images are directly determined by tear film thickness. RESULTS: As predicted by optical analysis of RI, the spatial distribution of gaps in the tear film seen with fluorescein appeared as pairs of light and dark contours in the RI images, and a precise correspondence between the spatial derivative of the FL image (slope) and the RI image was found. Both methods showed a gradual spreading of the tear disruption during blink suppression that varied tremendously among eyes in both time and spatial pattern. Resumption of normal blinking did not produce an immediate reconstitution of the normal tear film, and areas of tear break-up created during blink suppression remained abnormal for up to several minutes of normal blinking. CONCLUSIONS: Our analysis indicates that both FL and RI have the potential to quantify optical changes occurring during tear break-up. These results support an interpretation of RI as an intensity-based method for mapping the highly irregular optical aberrations of the eye produced by tear film disruption.

Adult↗

Energy and spatial distribution of multiple order Compton scatter in SPECT: a Monte Carlo investigation.

Energy and spatial projection distributions were simulated for gamma camera imaging of multiple order Compton scattered photons. SPECT imaging of a line source of radioactivity located in a water filled cylindrical phantom was modelled using Monte Carlo techniques. Photon trajectories were followed from emission to detection including the effects of all physical interactions and the resulting energy spectra and spatial projections were sorted as a function of the number of times the photon underwent Compton scattering before detection. Analysis of energy spectra demonstrates that Compton events up to second order overlap with the non-scattered events and distributions are peaked at lower energies as the scattering order increases. Analysis of spatial projections shows that, with increasing order, Compton events produce tails on the line spread function which progress from roughly exponential to nearly flat distributions. The use of Monte Carlo modelling thus allows a detailed investigation of the spatial and energy distribution of Compton scatter which could not be performed using present experimental techniques.

Models, Theoretical↗

Heterogeneity shapes invasion: host size and environment influence susceptibility to a nonnative pathogen.

Theoretical study of invasion dynamics has suggested that spatial heterogeneity should strongly influence the rate and extent of spreading organisms. However, empirical support for this prediction is scant, and the importance of understanding heterogeneity for real-world systems has remained ambiguous. This study quantified the influence of host and environmental heterogeneity on the dynamics of a 19-year disease invasion by the exotic and fatal pathogen, Phytophthora lateralis, within a stream population of its host tree, Port Orford cedar (Chamaecyparis lawsoniana). Using dendrochronology, we reconstructed the invasion history along a 1350-m length of infected stream, which serves as the only route of pathogen dispersal. Contrary to theoretical predictions, the temporal progression of the disease invasion was not related to a host's downstream spatial position, but instead was determined by two sources of heterogeneity: host size and proximity to the stream channel. These sources of heterogeneity influenced both the epidemic and endemic dynamics of this pathogen invasion. This analysis provides empirical support for the influence of heterogeneity on the invasion dynamics of a commercially important forest pathogen and highlights the need to incorporate such natural variability into both invasion theory and methods aimed at controlling future spread.

Animals↗

Temporal and spatial apodization in defocused acoustic transmission microscopy.

Two non-confocally adjusted spherical transducers are employed to implement an acoustic microscope operating in transmission with an approximately line-shaped point spread function (PSF). Such a PSF is of advantage in acoustic transmission line tomography and spatially resolved velocity measurements in solids. The foci of the transducers are viewed as diffraction-limited point transducers and appropriate time-selective signal acquisition is designed to restrict the ultrasound wave paths to the line connecting them. It is found that for typical commercially available transducers the largest contribution to the detected signal is not due to the direct ultrasound wave but due to the edge waves emanating from the rim of the focusing transducer. This poses constraints on achieving a line-shaped PSF in defocused acoustic transmission microscopy. It is shown that, due to the strong contribution from edge waves, it is impossible to achieve a line-shaped PSF in the case of application of a long exciting toneburst. The influence of the exciting pulse length, as well as the position of the time gate on the obtainable PSF is investigated.

Journal Article↗

Effects of axial spatial resolution and sampling on object detectability and contrast for multiplanar position emission tomography.

A multiplanar positron emission tomography (PET) system is simulated using Gaussian curves to model the axial point spread functions (PSFs) of the planes to study the effects of resolution and sampling. Poor spatial resolution or insufficient sampling may cause deleterious data losses or artifacts in the reconstructed image. For a multiplanar PET system with an axial full width at half-maximum (FWHM) of 6 mm and a 12 to 13 mm ring separation, a ripple in sensitivity of 9% is observed. A 1 mm object placed at the central direct plane results in detection of 59% of the signal in that plane. The theoretical observed contrast of a 3 mm object positioned at the center plane is 25% of the true contrast and decreases to 24% when the object is positioned between the central direct and cross planes. A PET system with an axial FWHM of 12 mm and a ring separation of 5-6 mm has a uniform sensitivity. A 1 mm object placed at the central direct plane detects 14% of the object signal in that plane. The theoretical observed contrast for a 3 mm object is 13% of true contrast when the object is positioned between the central direct and cross planes. It should be noted that all dimensions refer to the z direction through the center of the gantry in the simulated multiplanar system. The uniform sensitivity due to wider axial FWHMs decreases the amount of data loss for inter-ring gaps; however, the blurring associated with wider FWHMs decreases observed contrast.(ABSTRACT TRUNCATED AT 250 WORDS)

Health Physics↗

Potential role of nuclear magnetic resonance for the evaluation of trabecular bone quality.

This paper discusses two novel applications of nuclear magnetic resonance (NMR) as an investigational tool for the assessment of cancellous bone microarchitecture. It further outlines extensions of the method for in vivo clinical evaluation of bone strength in patients with skeletal disorders such as osteoporosis. The first method relies on the hypothesis that the presence of two phases of different magnetic permeability, i.e., bone and bone marrow, causes a spatial nonuniformity of the magnetic field across the measurement volume. The resulting spread in resonance frequency shortens the decay time constant (T2*) of the time domain proton signal in bone marrow or its substitute (water). Increased trabecular spacing, such as it occurs in osteoporosis, reduces the spatial field inhomogeneity and thus prolongs T2*, which has been shown both in vitro and in vivo. Subjects with osteoporosis, characterized by either low bone mineral density and/or spine compression fractures, have T2* values that are significantly prolonged. The second method focuses on a direct measurement of micromorphometric parameters of cancellous bone, using the principles of proton NMR microscopy in conjunction with computer processing of the resulting digital images. Image contrast between the trabeculae and the intertrabecular space is based on the marrow protons providing a signal, as opposed to bone, which appears with background intensity. Once tissues have been classified (into bone and marrow), for example, by means of a histogram-based segmentation algorithm, bone area fraction, mean trabecular plate density (MTPD), and mean trabecular plate thickness (MTPT) can be computed without the need for further operator intervention.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Pursuit-evasion predator-prey waves in two spatial dimensions.

We consider a spatially distributed population dynamics model with excitable predator-prey kinetics, where species propagate in space due to their taxis with respect to each other's gradient in addition to, or instead of, their diffusive spread. Earlier, we have described new phenomena in this model in one spatial dimension, not found in analogous systems without taxis: reflecting and self-splitting waves. Here we identify new phenomena in two spatial dimensions: unusual patterns of meander of spirals, partial reflection of waves, swelling wave tips, attachment of free wave ends to wave backs, and as a result, a novel mechanism of self-supporting complicated spatiotemporal activity, unknown in reaction-diffusion population models.

Adaptation, Physiological↗

Emergent trade-offs and selection for outbreak frequency in spatial epidemics.

Nonspatial theory on pathogen evolution generally predicts selection for maximal number of secondary infections, constrained only by supposed physiological trade-offs between pathogen infectiousness and virulence. Spread of diseases in human populations can, however, exhibit large scale patterns, underlining the need for spatially explicit approaches to pathogen evolution. Here, we show, in a spatial model where all pathogen traits are allowed to evolve independently, that evolutionary trajectories follow a single relationship between transmission and clearance. This trade-off relation is an emergent system property, as opposed to being a property of pathogen physiology, and maximizes outbreak frequency instead of the number of secondary infections. We conclude that spatial pattern formation in contact networks can act to link infectiousness and clearance during pathogen evolution in the absence of any physiological trade-off. Selection for outbreak frequency offers an explanation for the evolution of pathogens that cause mild but frequent infections.

Biological Evolution↗