Print Email Facebook Twitter Co-Orientation: Quantifying Simultaneous Co-Localization and Orientational Alignment of Filaments in Light Microscopy Title Co-Orientation: Quantifying Simultaneous Co-Localization and Orientational Alignment of Filaments in Light Microscopy Author Nieuwenhuizen, R.P.J. Nahidiazar, L. Manders, E.M.M. Jalink, K. Stallinga, S. Rieger, B. Faculty Applied Sciences Department ImPhys/Imaging Physics Date 2015-07-10 Abstract Co-localization analysis is a widely used tool to seek evidence for functional interactions between molecules in different color channels in microscopic images. Here we extend the basic co-localization analysis by including the orientations of the structures on which the molecules reside. We refer to the combination of co-localization of molecules and orientational alignment of the structures on which they reside as co-orientation. Because the orientation varies with the length scale at which it is evaluated, we consider this scale as a separate informative dimension in the analysis. Additionally we introduce a data driven method for testing the statistical significance of the co-orientation and provide a method for visualizing the local co-orientation strength in images. We demonstrate our methods on simulated localization microscopy data of filamentous structures, as well as experimental images of similar structures acquired with localization microscopy in different color channels. We also show that in cultured primary HUVEC endothelial cells, filaments of the intermediate filament vimentin run close to and parallel with microtubuli. In contrast, no co-orientation was found between keratin and actin filaments. Co-orientation between vimentin and tubulin was also observed in an endothelial cell line, albeit to a lesser extent, but not in 3T3 fibroblasts. These data therefore suggest that microtubuli functionally interact with the vimentin network in a cell-type specific manner. Subject vimentintubulinskeratinsendothelial cellsfibroblastscytoskeletonfluorescence microscopyFourier analysisOA-Fund TU Delft To reference this document use: http://resolver.tudelft.nl/uuid:b9548433-b619-435a-807e-a86289c218c6 Publisher Public Library of Science ISSN 1932-6203 Source https://doi.org/10.1371/journal.pone.0131756 Source PLoS ONE, 10 (7), 2015 Part of collection Institutional Repository Document type journal article Rights © 2015 The Author(s)This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Files PDF Rieger_2015.pdf 9.37 MB Close viewer /islandora/object/uuid:b9548433-b619-435a-807e-a86289c218c6/datastream/OBJ/view