Timelapse imaging, boundary segmentation, and cell tracking data from cultured proliferating Drosophila wing disc epithelia

Overview This dataset includes Drosophila wing imaginal disc epithelial tissues cultured ex vivo while imaging of Ecadherin-GFP (to mark adherens junctions on the membrane) with spinning disc confocal microscopy. Individual cells of the pouch region of the tissue were segmented and tracked over the movie, and a database was created to house relevant cell parameters using TissueMiner (Etournay et al 2016). Data were generated at the Light Microscopy Facility of the Max Planck Institute for Molecular Cell Biology, Dresden, Germany by Natalie Dye from 2014-2016. This work was supported by core funding from the Max Planck Society, as well as DFG grants to S. Eaton (EA4/10-1, EA4/10-2). Methods Experimental methods are described in Dye et al 2017 and Dye et al 2021. In brief, mid-third instar wing discs were cultured ex vivo in Grace's insect media supplemented with 5% FBS and 20nM 20-hydroxyecdysone. Tissues expressed a knock-in GFP-tag of E-cadherin to mark apical adherens juncitons. Spinning disc confocal was used for imaging Z-stacks (0.5um step size) at 5min intervals. Stacks were flipped if necessary to have the anterior of the tissue on the left. The apical surface was then projected to 2D using a custom surface projection algorithm (see Dye et al 2017), with the corresponding heightmap used to measure local tissue curvature and adjust for distortions to cell area and elongation due to projection. Cell segmentation and tracking was performed with TissueAnalyzer (Fiji plugin, Aigouy et al 2016) with manual correction. TissueMiner (Etournay et al 2016) was then used to generate a relational database containing cell area/elongation, lineage, position, etc. Contents Each zip folder contains a different tissue - included are 5 independent biological replicates. They are named: Ecd_20141010_P2 (Movie 1 in Dye et al 2017 and 2021) Ecd_20150310_P2 (Movie 2 in Dye et al 2017 and 2021) Ecd_20150418_P1 (Movie 3 in Dye et al 2017 and 2021) Ecd_20150608_P1 (Movie 4 in Dye et al 2021)Ecd_20161213_F0 (Movie 5 in Dye et al 2021) XY pixel size: for Movies 1-4: 0.208x0.208um; for Movie 5: 0.196x0.196um Each movie folder contains a database file (sqlite) generated by TissueMiner (Etournay et al 2016) and tables of height-corrected cell areas and height-corrected triangle elongations (accounting for errors due to tissue curvature - see Dye et al 2021). Also included is a subfolder called "Segmentation". Within that subfolder are the individual timepoint images and a subfolder for each timepoint containing the segmentation data for that timepoint (a subset of files generated upon segmentation with the FIJI plugin TissueAnalyzer - Aigouy et al 2016). The exact time intervals are recorded in the file "cumultimesec.txt" file - which gives the actual time (in sec) when the Z-stack finished imaging (cumulative from start). The transformation.txt file is used to orient the tissue and database such that anterior is on the left and dorsal up. Finally the dataset also includes the center positions for each movie calculated from cell elongation patterns (see Dye et al 2021). **Please note that the movie "Ecd_20150310_P2" (Movie 2 in Dye et al 2017 and 2021) is slightly abberant. We have noted a region near the dorsal-ventral boundary on the anterior side where there is abnormally high amounts of cell extrusions and a lack of cell proliferation. Consider excluding this movie, depending on your use case.