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Image cytometry has emerged as a valuable in vitro screening tool and advances in automated microscopy have made it possible to readily analyze large cellular populations of image data. The purpose of this paper is to illustrate the viability of using cell shape to test equality of cell populations based on image data. Shape space theory is reviewed, from which differences between shapes can be quantified in terms of geodesic distance. Several multivariate nonparametric statistical hypothesis tests are adapted to test equality of cell populations. It is illustrated that geodesic distance can be a better feature than cell spread area and roundness in distinguishing between cell populations. Tests based on geodesic distance are able to detect natural perturbations of cells, whereas Kolmogorov-Smirnov tests based on area and roundness are not.

Cell segmentation is a critical step in the analysis pipeline for most imaging cytometry experiments and evaluating the performance of segmentation algorithms is important for aiding the selection of segmentation algorithms. Four popular algorithms are evaluated based on their cell segmentation performance. Because segmentation involves the classification of pixels belonging to regions within the cell or belonging to background, these algorithms are evaluated based on their total misclassification error. Misclassification error is particularly relevant in the analysis of quantitative descriptors of cell morphology involving pixel counts, such as projected area, aspect ratio and diameter. Since the cumulative distribution function captures completely the stochastic properties of a population of misclassification errors it is used to compare segmentation performance.