DOI: 10.6084/m9.figshare.26346454
title: Tumour-stroma_spheroid_multicultures_features

description: ARTICLE (when using these files, please, cite the following article):Akos Diosdi, Filippo Piccinini, Timea Boroczky, Gabriella Dobra, Gastone Castellani, Krisztina Buzas, Peter Horvath, Maria Harmati. "Single-cell light-sheet fluorescence 3D images of tumour-stroma spheroid multicultures". (2024)DESCRIPTION OF THE FILES:The dataset contains 90 multi-tiff fluorescence images of control and drug-treated multiculture 3D tumour models taken 24, 48, and 96 hours after seeding. Each spheroid model includes one tumour cell line (either T-47D ductal carcinoma, A375 melanoma, or MG-63 osteosarcoma) and two stromal cell lines, <i>i.e.</i> MRC-5 fibroblasts, and EA.hy926 endothelial cells (ATCC, Manassas, Virginia, USA). Before seeding, each cell line was stained differently with CellTracker dyes based on the manufacturer’s instructions (Invitrogen, Thermo Fisher Scientific), precisely MRC-5 - Deep Red, EA.hy926 - Green CMFDA, and tumour cell lines - Orange CMTMR dyes. Additionally, each tumour model was stained with Hoechst 33342 in DMEM for 60 min before imaging. Overall, 4 channels were acquired: Nuclei - 405 (ch001), EA.hy926 - 488 (ch002), tumour - 552 (ch003), and MRC-5 - 638 (ch004) with the Leica True Confocal Scanning (TCS) SP8 Digital LightSheet (DLS) microscope. The multi-page 16-bit Tiff files have a resolution of 2048 × 2048, with pixel size 0.14370117 µm, and the distance between each image in each z-stack is 3.7 µm. Additionally, Maximum Intensity Projection (MIP) images were generated and morphological features were extracted from every spheroid in the collection using a software tool known as AnaSP. The extracted features of the 90 spheroids are included.FILE NAMES:All the files in this collection follow this terminology:<br>“TumourType_+*_TimePoint_SpheroidID_channelID.tif”*If treatment was applied.MODELS:A: T-47D<br>A+: T-47D treated with 0.6 µM doxorubicin<br>B: A375<br>B+: A375 treated with 0.6 µM doxorubicin<br>C: MG-63<br>C+: MG-63 treated with 0.6 µM doxorubicin<br>TIME POINTS:24h<br>48h<br>96h<br>For each tumour cell line and time point, five spheroids have been imaged under both drug-treated and control conditions.MAXIMUM INTENSITY PROJECTION IMAGES:Maximum Intensity Projection (MIP) images were created by combining all the z-stack images of spheroids. Channels were separated and the merged images for three channels (merge1: 488, 552, and 638) and four channels (merge_all: 405, 488, 552, and 638) were included in the collection.IMAGE ANALYSIS:Using a software called AnaSP, morphological features have been extracted from each spheroid reported in the collection. A table containing the extracted features of the 90 spheroids is included.<br><br>Features_all.xls<br>Features_pixel.csv<br>Features_um.csv<br>MAIN CONTACTS:Maria Harmati, Synthetic and Systems Biology Unit, HUN-REN Biological Research Centre (HUN-REN BRC), 6726 Szeged, Hungary<br>Email: harmati.maria@brc.hu<br>COPYRIGHT:* Copyright (c) 2024, Akos Diosdi, Peter Horvath, Maria Harmati,* HUN-REN Biological Research Centre (HUN-REN BRC) HUN-REN, Szeged, Hungary* All rights reserved.** Redistribution and use of the material, with or without modification, is provided for academic research purposes only.** This material is free; you can redistribute it and/or modify it under the terms of the CC BY 4.0.