C.M. RUSU*#, MIHAELA BACALUM*, ANTONIA TEONA DEFTU**, BEATRICE MIHAELA RADU**, M. RADU*
*Department of Life and Environmental Physics, “Horia Hulubei” National Institute of Physics and Nuclear Engineering, 30, Reactorului st, Măgurele, Romania
**Department of Anatomy, Animal Physiology and Biophysics, Faculty of Biology, University of Bucharest, 91–95, Splaiul Independenței, Bucharest, Romania
The actin cytoskeleton is a basic determinant of cellular morphology and motility. Actin cytoskeleton study is important because a wide spectrum of cellular processes can be monitored through the conformational changes that occur at its level. A template-based method for linear detection was used to process and perform a quantitative analysis of cytoskeleton morphology in various cell types. This approach is fitted for a precise calculation of various parameters such as fiber length and cytoskeleton polarization, and can be used to follow the changes in the morphology of the actin cytoskeleton due to physiological modifications. In our study, several images of four different cell lines: bEnd.3, BJ, MG-63 and OLN-93 were analyzed in control conditions, in order to assess the structural organization of the actin cytoskeleton. A comparative analysis was also performed on the robustness and adaptability of the proposed algorithm, using the images of the four cell lines. The aim of this study was to establish how well the program recognizes the fibers apart from other cellular structures and debris. Following the analysis, criteria were set for the adjustment of computational parameters (e.g., angular resolution, kernel matrix size, etc.). The use of the segmentation algorithm provided consistent quantitative information, revealing the distributions of important morphological features. The adaptive character of the method, due to the manipulation of computational parameters, enables an efficient segmentation of the microscopy images. Thus, a high precision quantitative analysis of the microscopy images was performed, for the assessment fiber morphology and determination of the optimal set of computational parameters used for the digital extraction of the fibers.
Key words: cytoskeleton, actin, microscopy, confocal, digital, analysis.
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