Image processing and analysis of fluorescent labelled cytoskeleton

Cytoskeleton elements play an important role in cellular processes includin g normal and pathological conditions or even cell death. Although immunoflu orescence microscopy techniques have proved useful to study the cytoskeleto n, description and comparative results commonly rely on the direct visual a nalysis 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 c omponents. Different structural quantifiable characteristics using TPA faci lities (densitometric profile, number of branched points, Euler number, con vexity, orientation value) allow accurate characterization and differentiat ion of the three types of fibre nets studied (microtubules, actin microfila ments, and cytokeratin intermediate filaments). The application of the anal ytical methods presented here provides information concerning the organizat ion and modification oi cytoskeleton components, and could aid in the furth er understanding of structural and functional aspects of the cytoskeleton u nder normal and pathological conditions. (C) 1999 Elsevier Science Ltd.