Image processing and analysis of fluorescent labelled cytoskeleton

Cytoskeleton elements play an important role in cellular processes including normal and pathological conditions or even cell death. Although immunofluorescence microscopy techniques have proved useful to study the cytoskeleton, description and comparative results commonly rely on the direct visual analysis of the targeted cells. We report here an objective and quantitative method to analyze the cytoskeletal pattern of cultured HeLa cells based on image processing and analysis (IPA) of fluorescent labelled cytoskeleton components. Different structural quantifiable characteristics using IPA facilities (densitometric profile, number of branched points, Euler number, convexity, orientation value) allow accurate characterization and differentiation of the three types of fibre nets studied (microtubules, actin microfilament, and cytokeratin intermediate filaments). The application of the analytical methods presented here provides information concerning the organization and modification of cytoskeleton components, and could aid in the further understanding of structural and functional aspects of the cytoskeleton under normal and pathological conditions.