a numerical analysis model for interpretation of flow cytometric studies of ex vivo phagocytosis数值分析模型的解释流仪研究体外吞噬作用.pdfVIP

A Numerical Analysis Model for Interpretation of Flow Cytometric Studies of Ex Vivo Phagocytosis 1,2 3 2 Ted S. Strom *, Praveen Anur , Amanda Prislovsky 1 Department of Pathology and Laboratory Medicine, University of Tennessee Health Sciences Center, Memphis, Tennessee, United States of America, 2 Department of Pathology and Laboratory Medicine, Memphis Veterans Administration Medical Center, Memphis, Tennessee, United States of America, 3 Oregon Health Sciences University, Portland, Oregon, United States of America Abstract The study of ex vivo phagocytosis via flow cytometry requires that one distinguish experimentally between uptake and adsorption of fluorescently labeled targets by phagocytes. Removal of the latter quantity from the analysis is the most common means of analyzing such data. Because the probability of phagocytosis is a function of the probability of adsorption, and because partially quenched fluorescence after uptake often overlaps with that of negative controls, this approach is suboptimal at best. Here, we describe a numerical analysis model which overcomes these limitations. We posit that the random adsorption of targets to macrophages, and subsequent phagocytosis, is a function of three parameters: the ratio of targets to macrophages (m), the mean fluorescence intensity imparted to the phagocyte by the internalized target (alpha), and the probability of phagocytosis per adsorbed target (p). The potential values of these parameters define a parameter space and their values at any point in parameter space can be used to predict the fraction of adsorption(+) and [adsorption(2), phagocytosis(+)] cells that might be observed experimentally. By systematically evaluating th