The latest version of Grafeo (v3) incorporates development for the following article: "Multicolor 3D-dSTORM Reveals Native-State Ultrastructure of Polysaccharides' Network during Plant Cell Wall Assembly" iScience 2020, https://doi.org/10.1016/j.isci.2020.101862. The updated manual and sample 3-color 3D dSTORM data will be published shorrtly. Grafeo is a Matlab based program for the analysis of single-molecule localization microscopy (SMLM) data e.g., dSTORM or PALM. It works on a single molecule coordinates rather than pixelated images. Grafeo uses various pointillist approaches previously described in the literature, e.g., Voronoi Tessellation, Delaunay Triangulation, Ripley’s function. Grafeo was first developed for the following publication (version 1): Haas et al., "Single-molecule localization microscopy reveals molecular transactions during RAD51 filament assembly at cellular DNA damage sites", Haas et. Al., NAR 2018, https://doi.org/10.1093/nar/gkx1303. Grafeo version 2.1.0 was developed for the following publication: “Pectin homogalacturonan nanofilament expansion drives morphogenesis in plant epidermal cells”, Haas et al., Science 2020. DOI: 10.1126/science.aaz5103 The current version 2.2.0 includes new features, such as three-color data visualization and processing, rapid calculation of 3D boundary corrected Ripley's and PCF function for small ROIs, and others. The 2.2.0 updates were developed for the following article, currently under review in iScience: “Multicolor 3D-dSTORM reveals native-state ultrastructure of polysaccharides’ network during plant cell wall assembly”, Peaucelle et. al. If you have any questions or comments, please contact the author via email: [email protected], [email protected], or Twitter @KalinaHaas. You can download an example of analyzed data here: http://dx.doi.org/10.17632/z53y4654fs.1.