Dynamics of Cellular Responses studied by Quantitative Colocalization Analysis
Quantitative colocalization analysis is a tool to objectively estimate the degree of overlap (colocalization) of fluorescently labelled antigens in biological microscopy. It allows the drawing of unbiased conclusions about their interaction and function. This article summarizes the theoretical basis of quantitative colocalization analysis and describes the use of the technique. The applicability and correct interpretation of the results of colocalization coefficients calculations is given for the investigation of the role of platelet activating factor (PAF) and its receptor (PAF-R) in the recruitment of eosinophils into the conjunctiva. Quantitative colocalization analysis helped to determine that the recruitment of eosinophils was accomplished by direct action of PAF on the PAF-R and that the interaction of both PAF-R and major basic protein antigens with PAF was required for eosinophil influx.
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