0000000000072837
AUTHOR
M. E. Díaz
Fuzzy temporal random sets with an application to cell biology
Total Internal Reflection Fluorescence Microscopy (TIRFM) greatly facilitates to imaging the first steps of endocytosis, a process whereby cells traffic cargo from the cell surface to endosomes. Using TIRFM, fluorescent-tagged endocytic proteins are observed as overlapped areas forming random clumps of different sizes, shapes and durations. A common procedure to segment these objects consists of thresholding the original gray-level images to produce binary sequences in which a pixel is covered or not by a given fluorescent-tagged protein. This binary logic is not appropriate because it leaves a free tuning parameter to be set by the user which can influence on the conclusions of the statist…
Experimental Analysis of the Multipactor Effect With RF Pulsed Signals
The main goal of this letter is the analysis of the multipactor effect within a coaxial waveguide excited by an RF pulsed signal. The variation of the multipactor RF voltage thresholdwiththe ON interval length of the pulse has been analyzed. To reach this aim, an in-house multipactor simulation code based on the Monte-Carlo algorithm has been implemented. The numerical simulations show that the multipactor RF voltage threshold increases as the ON pulse interval diminishes. In addition, an experiment was carried out to validate the proposed theoretical model, and demonstrating the excellent agreement between the theory and the experimental data. Finally, the results are compared with the 20-…
Measuring Spatiotemporal Dependencies in Bivariate Temporal Random Sets with Applications to Cell Biology
Analyzing spatiotemporal dependencies between different types of events is highly relevant to many biological phenomena (e.g., signaling and trafficking), especially as advances in probes and microscopy have facilitated the imaging of dynamic processes in living cells. For many types of events, the segmented areas can overlap spatially and temporally, forming random clumps. In this paper, we model the binary image sequences of two different event types as a realization of a bivariate temporal random set and propose a nonparametric approach to quantify spatial and spatiotemporal interrelations using the pair correlation, cross-covariance, and the Ripley K functions. Based on these summary st…
Measuring the Spatial Homogeneity in Corneal Endotheliums by Means of a Randomization Test
Quantification of regularity of cell sizes and the spatial arrangement of cells in corneal endotheliums becomes of a great importance associated to stress situations such as cataract surgery, corneal transplantation or implantation of intra-ocular lenses. A new index of regularity of the spatial distribution of cell sizes in corneal endotheliums is proposed. The corneal endothelium is described by means of a spatial marked point pattern (the cell centroids marked with the cell areas). The hypothesis of no dependency between mark and locations is tested by a Monte Carlo test. The new index is the p-value of the test validating the hypothesis. Pairs of endotheliums from different eyes of the …
Accurate detection and characterization of corner points using circular statistics and fuzzy clustering
Accurate detection and characterization of corner points in grey level images is considered as a pattern recognition problem. The method considers circular statistic tests to detect 2D features. A fuzzy clustering algorithm is applied to the edge orientations near the prospective corners to detect and classify them. The method is based on formulating hypotheses about the distribution of these orientations around an edge, corner or other 2-D feature. The method may provide accurate estimates of the direction of the edges that converge in a corner, along with their confidence intervals. Experimental results show the method to be robust enough against noise and contrast changes. Fuzzy membersh…
Studying endocytosis in space and time by means of temporal Boolean models
Endocytosis is a process by which cells carry traffic from the extracellular space into various intracellular compartments. Visualization of fluorescently tagged clathrin proteins (mediators of endocytosis) allows us to image endocytosis in real time. When imaging the plasma membrane, areas of fluorescence generated by different endocytic processes overlap spatially and temporally, forming random clumps. Here, a sequence of segmented clathrin spots is considered a realization of a non-isotropic 3D Boolean model. Estimates of the intensity, the mean perimeter and the density function of the durations of endocytic events are obtained.
Analyzing Protein-Protein Spatial-Temporal Dependencies from Image Sequences Using Fuzzy Temporal Random Sets
Total Internal Reflection Fluorescence Microscopy (TIRFM) allows us to image fluorescenttagged proteins near the plasma membrane of living cells with high spatial-temporal resolution. Using TIRFM imaging of GFP-tagged clathrin endocytic proteins, areas of fluorescence are observed as overlapping spots of different sizes and durations. Standard procedures to measure protein-protein colocalization of dual labeled samples threshold the original graylevel images to segment areas covered by different proteins. This binary logic is not appropriate as it leaves a free tuning parameter which can influence the conclusions. Moreover, these procedures rely on simple statistical analysis based on corre…
Analysis of Spatially and Temporally Overlapping Events with Application to Image Sequences
Counting spatially and temporally overlapping events in image sequences and estimating their shape-size and duration features are important issues in some applications. We propose a stochastic model, a particular case of the nonisotropic 3D Boolean model, for performing this analysis: the temporal Boolean model. Some probabilistic properties are derived and a methodology for parameter estimation from time-lapse image sequences is proposed using an explicit treatment of the temporal dimension. We estimate the mean number of germs per unit area and time, the mean grain size and the duration distribution. A wide simulation study in order to assess the proposed estimators showed promising resul…
Phosphatidylinositol phosphate kinase type I gamma regulates dynamics of large dense-core vesicle fusion.
Phosphatidylinositol-4,5-bisphosphate was proposed to be an important regulator of large dense-core vesicle exocytosis from neuroendocrine tissues. Here, we have examined the kinetics of secretion in chromaffin cells from mice lacking phosphatidylinositol phosphate kinase type Iγ, the major neuronal phosphatidylinositol-4-phosphate 5-kinase. Absence of this enzyme caused a reduction of the readily releasable vesicle pool and its refilling rate, with a small increase in morphologically docked vesicles, indicating a defect in vesicle priming. Furthermore, amperometry revealed a delay in fusion pore expansion. These results provide direct genetic evidence for a key role of phosphatidylinositol…
Set Descriptors for Visual Evaluation of Human Corneal Endothelia
Images of corneal endothelium obtained from specular microscopy are of great importance in the evaluation of the corneal endothelium status. Several commercial tools provide some numerical descriptors to characterize these images in terms of cell density, hexagonality, and some descriptive statistics of the cell areas. However, it is a too simple analysis that only detects severe abnormal endothelia with many irregular and large cells. Detection of subtle abnormalities needs a more refined analysis. This paper proposes a shape-size descriptor based on some modified versions of the geometric covariogram. This descriptor is presented as a valid alternative to the classical analysis that provi…
Spatio-Temporal Analysis of Constitutive Exocytosis in Epithelial Cells
Exocytosis is an essential cellular trafficking process integral to the proper distribution and function of a plethora of molecules, including transporters, receptors, and enzymes. Moreover, incorrect protein targeting can lead to pathological conditions. Recently, the application of evanescent wave microscopy has allowed us to image the final steps of exocytosis. However, spatio-temporal analysis of fusion of constitutive vesicular traffic with the plasma membrane has not been systematically performed. Also, the spatial sites and times of vesicle fusion have not yet been analyzed together. In addition, more formal tests are required in testing biological hypotheses, rather than visual insp…
Bayesian estimation of edge orientations in junctions
Abstract Junctions, defined as those points of an image where two or more edges meet, play a significant role in many computer vision applications. Junction detection is a widely treated problem, and some detectors can provide even the directions of the edges that meet in a junction. The main objective of this paper is the precise estimation of such directions. It is supposed that the junction point has been previously found by some detector. Also, it is assumed that samples, possibly noisy, of orientations of the edges found in a circular window surrounding the point are available. A mixture of von Mises distributions is assumed for these data, and then a Bayesian methodology is applied to…
Granulometric moments and corneal endothelium status
Abstract Specular microscopy is a common practice in Ophthalmology. The corneal endothelium status is usually evaluated by means of the density, the hexagonality, the mean, the standard deviation and the coefficient of variation of cell areas. We propose to replace the cell area moments by the corresponding moments of a different probability distribution, the granulometric size distribution associated to a disc. All cells touching the frame are ignored by the area moments but used by the granulometric moments. Twenty images have been analyzed. When the size of the focused region is reduced, the area moments show a greater variation than the corresponding granulometric moments.
MORPHOMETRIC ANALYSIS OF HUMAN CORNEAL ENDOTHELIUM BY MEANS OF SPATIAL POINT PATTERNS
This paper presents a method for detecting abnormalities in spatial arrangements of cells within any tissue that can be described by different sets of relevant points. The method has been applied to the detection of subtle abnormalities in corneal endothelia. Images of this type of tissue can be characterized by two types of points: cell centroids and triple points associated with the apical intersections as it was proposed by Díaz.7 Both types of points jointly considered are modeled using a bivariate spatial point process; then a statistical analysis based on certain distributional descriptors proposed by Doguwa4,9 is carried out to discriminate severe and subtle abnormalities from contr…
Multipactor RF Breakdown in Coaxial Transmission Lines With Digitally Modulated Signals
The aim of this paper is the study of the RF multipactor breakdown in coaxial transmission lines excited by a single carrier with a digitally modulated signal. Employing an in-house developed code, numerical simulations are performed to determine the RF multipactor voltage threshold for several digitally modulated signals under different modulations schemes: quadrature phase-shift keying, 16-quadrature amplitude modulation, 16-amplitude and phase-shift keying, and 32-amplitude and phase-shift keying. Moreover, a coarse method based on the envelope integration to determine the RF multipactor voltage threshold when involving arbitrary digital modulations is also presented. These results are a…
A gray-level 2D feature detector using circular statistics
Abstract This paper presents a new method for corner and circular feature detection in gray-level images. It is based on the application of standard statistical techniques to the distribution of gradient orientations in a circular neighborhood of the prospective feature point. An evaluation using standard procedures and a comparison with other approaches is presented. Results show the robustness of this method as compared to the other corner detectors analyzed. The main novelties are the possibility of detecting points that are centers of circular symmetries, and discriminating between junctions, which are classified into corners (two-edge junctions) and multiple edge junctions.
Testing abnormality in the spatial arrangement of cells in the corneal endothelium using spatial point processes
The study of central corneal endothelium morphology is important in Ophthalmology. Some of the pathologies that could compromise endothelial cell morphology are trauma, cataract, surgery, use of contact lenses, corneal dystrophies or degenerations. The quantitative analysis of cell shape and cellular pattern is more sensitive in detecting subtle changes in endothelial morphology than cell density measurement or cell area analysis. In this paper, the morphology of the central cornea, the most important area from the point of view of vision, is studied through an associated bivariate spatial point pattern: the centroids of the cells and the triple points, that is, the points where three diffe…
Novel multipactor studies in RF satellite payloads: Single-carrier digital modulated signals and ferrite materials
In this work it is reviewed the most novel advances in the multipactor RF breakdown risk assessment devoted to RF satellite microwave passive devices employed in space telecommunication systems. On one side, it is studied the effect of transmitting a single-carrier digital modulated signal in the multipactor RF voltage threshold in a coaxial line. On the other hand, an analysis of the multipactor phenomenon in a parallel-plate waveguide containing a magnetized ferrite slab it is presented.
Granulometric analysis of corneal endothelium specular images by using a germ–grain model
Specular microscopy is widely used to study the human corneal endothelium status in vivo. In this paper, the corneal endothelium is represented as a binary image composed of the cell inscribed circles. The granulometric distribution function of the complement of this image is used as a functional descriptor, which provides information about the shape, size and spatial arrangement of cells. Experimental evaluation using bootstrap techniques shows its ability to discriminate between controls and pathological cases. It represents a reliable and graphical alternative to the classical indices (cell density, hexagonality and coefficient of variation of cell areas), which behave poorly when detect…