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P8010
Quantitative Analysis of Angiogenesis & Cytoskeletal Formation with IN Cell Investigator
Presenter Presenter Ian Davies, GE Healthcare, United Kingdom
Additional Authors: A. Zaltsman, C. Hather, Y. Alexandrov, A. Kuzmin, A. Santos, R. Martin, L. Roquemore, I. Davies
Angiogenesis and filamentous actin formation are examples of cell processes that present unique challenges for quantitative image analysis, in terms of spatial distribution of structures in 2- or 3-  dimensions and the need to detect elongated shapes.  To address such challenges, we have developed new features and Context Modules for IN Cell Investigator image analysis software.  A new extended focus transformation tool allows generation of a 2-D composite projection from an image stack acquired in 3 dimensions (3-D z-series). This feature is utilized for analysis of angiogenesis formation from cells cultured on a 3-D matrix. We have also developed an Angiogenesis Context Module to enable high-throughput, high-content analysis of screens for pro- and anti-angiogenic compounds.  The Angiogenesis Context Module is designed for analysis of angiogenesis in 96-well plates containing a specially formulated fibrin matrix, and employs a new shape selector segmentation feature to assist in identification of elongated multi-cellular or macromolecular formations. After application of the extended focus transformation, a composite image stack can be analyzed with the context module to characterize development of angiogenic structures.   The new shape selector segmentation feature was also utilized for the analysis of F-actin fiber reorganization in HeLa, CHO and NIH 3T3 cells in response to cytochalasin D or staurosporine treatments. The assay characterizes disruption of actin polymerization or reorganization of the actin cytoskeleton network and is suitable for screening, producing Z’ values of 0.55 and 0.7 for HeLa and CHO cells respectively.