The analysis of fluorescence microscopy of cells often requires the determination of cell edges. This is typically done using segmentation techniques that separate the cell objects in an image from the surrounding background. This study compares segmentation results from nine different segmentation techniques applied to two different cell lines and five different sets of imaging conditions. Significant variability in the results of segmentation was observed that was due solely to differences in imaging conditions or applications of different algorithms. We quantified and compared the results with a novel bivariate similarity index metric that evaluates the degree of underestimating or overestimating a cell object. The results show that commonly used threshold-based segmentation techniques are less accurate than k-means clustering with multiple clusters. Segmentation accuracy varies with imaging conditions that determine the sharpness of cell edges and with geometric features of a cell. Based on this observation, we propose a method that quantifies cell edge character to provide an estimate of how accurately an algorithm will perform. The results of this study will assist the development of criteria for evaluating interlaboratory comparability. Published 2011 Wiley-Liss, Inc.y Key terms fluorescence microscopy; k-means cluster; image segmentation; cell edge; bivariate similarity index NUMEROUS areas of biomedical research rely on imaging of cells to provide information about molecular and phenotypic responses of cells to pharmaceuticals, toxins, and other environmental factors (1,2). Cell imaging is widely used in biological experiments because it can provide information on several relevant scales simultaneously. Molecular and supramolecular scales can be probed with the use of antibody and other specific affinity reagents and with fluorescent proteins. Gross phenotypic characteristics of cells that characterize their ultimate functional state can be examined in the same experiment, and often the temporal regime can be probed simultaneously. Together, these applications of cell imaging allow inference about the molecular details and the complex outcomes of the cellular biochemistry.Because of the enormous number of parameters that may influence a biological outcome, cell imaging experiments are often done in a ''high content '' mode (3,4), where large numbers of paired and replicate experiments are carried out simultaneously and result in very large (often gigabyte) image datasets. Such a large volume of image data precludes visual inspection of every image, and automated image processing and analysis is the only viable approach to data analysis.Segmentation of cell objects is a common image analysis operation that provides spatial and other features of identified objects and often precedes other operations to quantify parameters such as intracellular fluorescence. Segmentation can pose significant challenges to automated image processing and analysis. Because morphological features are often important in...

Analysis of lineaments from satellite images normally includes the determination of their orientation and density. The spatial variation in the orientation and/or number of lineaments must be obtained by means of a network of cells, the lineaments included in each cell being analysed separately. The program presented in this work, LINDENS, allows the density of lineaments (number of lineaments per km 2 and length of lineaments per km 2 ) to be estimated. It also provides a tool for classifying the lineaments contained in dierent cells, so that their orientation can be represented in frequency histograms and/or rose diagrams. The input ®le must contain the planar coordinates of the beginning and end of each lineament. The density analysis is done by creating a network of square cells, and counting the number of lineaments that are contained within each cell, that have one of their ends within the cell or that cross-cut the cell boundary. The lengths of lineaments are then calculated. To obtain a representative density map the cell size must be ®xed according to: (1) the average lineament length; (2) the distance between the lineaments; and (3) the boundaries of zones with low densities due to lithology or outcrop features. An example from the Neogene Duero Basin (Northern Spain) is provided to test the reliability of the density maps obtained with dierent cell sizes. 7

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