Dataset title: UT-GPCR003 Fluorescence anisotropy and microscopy measurements experimental metadata of ligand binding to M4 muscarinic receptors Authors: Tahk, Maris-Johanna; Torp, Jane; Ali, Mohammed A.S.; Fishman, Dmytro; Parts, Leopold; Grätz, Lukas; Müller, Christoph; Keller, Max; Veiksina, Santa; Laasfeld, Tõnis; Rinken, Ago Contact information: laasfeld@ut.ee; ago.rinken@ut.ee Chair of Bioorganic chemistry, Institute of Chemistry, University of Tartu General dataset description: The dataset was gathered as a part of Tahk MJ, Torp J, Ali MA, Fishman D, Parts L, Grätz L, Müller C, Keller M, Veiksina S, Laasfeld T, Rinken A. Live-cell microscopy or fluorescence anisotropy with budded baculoviruses-which way to go with measuring ligand binding to M4 muscarinic receptors?. bioRxiv. 2021 Jan 1. https://doi.org/10.1101/2021.12.22.473643 All data is provided two compressed zip folders: ASCII files and MIDAS files. ASCII files ASCII files are organized into four folders by experiment type (competition or saturation) Each file contains the raw measurement data from a single fluorescence anisotropy experiment and by fluorescence ligand used (UR-CG072 and UR-MK342). Each file is named as Date_Time_Ligand_Receptor-source_experiment type Date is given in the yymmdd format, time is given in hhmmss format MIDAS files MIDAS files are CSV or XLS files organized into two folders (Fluorescence anisotropy and Microscopy) Fluorescence anisotropy folder structure and naming conventions match the ASCII file organzation MIDAS files have a "MD-" prefix to differentiate them from other kinds of CSV and XLS files. MIDAS files integrate experimental measurement data and minimal necessary experiment metadata. More information about MIDAS files can be found from: doi: 10.1093/bioinformatics/btn018 and https://gpcr.ut.ee/uploads/ApareciumHelp/Aparecium.html?Welcome1.html by experiment type (competition or saturation) Methodologial information: All raw ASCII data has been gathered using SynergyNeo multimode plate reader calibrated as described in Thompson, R. B., Gryczynski, I., & Malicka, J. (2002). Fluorescence polarization standards for high-throughput screening and imaging. BioTechniques, 32(1), 34-42. Each raw ASCII file contains detailed metadata about specific acquisitionconditions in the header. The MIDAS were generated as described in the connected publication (https://doi.org/10.1101/2021.12.22.473643). Aparecium software was used for generating and analyzing the ASCII files available at https://github.com/laasfeld/Aparecium and https://gpcr.ut.ee/aparecium.html Data specific information: MIDAS file headers: Identifier columns (beginning with ID:) Treatment columns (beginning with TR:) M4R-BBV_ : BBVs with M4 muscarinic receptor on the surface UR-CG072 - Fluorescence ligand as defined in the accompanying publication UR-MK342 - Fluorescence ligand as defined in the accompanying publication Scopolamine, Atropine, UNSW-MK259, Carbachol, Pirenzepine, Pilocarpine, Acetylcholine, Arecoline, UR-SK75, UR-SK59 - competitive ligands as defined in the accompanying publication CHO-hM4 - CHO-K1-hM4R cell line as defined in the accompanying publication Units for each treatment are given after the last underscore in the column header. Time column (beginning with DA:) DA:ALL - time point for all measurments in the same row. Time is measured from addition of BBV for FA experiments and addition of ligands for microscopy experiments. Always measured in seconds. Measurement columns (beginning with DV:) In fluorescence anisotropy experiments: Parallel - Fluorescence intensity parallel to the excitation plane, arbitrary units Perpendicular - Fluorescence intensity perpendicular to the excitation plane, arbitrary units In microscopy expeirments: averageCellIntensity - average fluorescence channel pixel intensity of all pixels per well identified as cells by segmentation algorithm averageSecondaryImageIntensity - average fluorescence intensity of the fluorescence channel images averageNonCellIntensity - average fluorescence channel pixel intensity of all pixels per well identified as not cells by segmentation algorithm firstNonCellQuadrileIntensity - average fluorescence channel pixel intensity of the lowest 25% intensity pixels per well identified as not cells by segmentation algorithm averageUnmaskedCellIntensity - average fluorescence channel pixel intensity of all pixels per well identified as cells by segmentation algorithm without considering the quality mask averageUnmaskedNonCellIntensity - average fluorescence channel pixel intensity of all pixels per well identified as not cells by segmentation algorithm without considering the quality mask firstUnmaskedNonCellQuadrileIntensity - average fluorescence channel pixel intensity of the lowest 25% intensity pixels per well identified as not cells by segmentation algorithm without considering the quality mask averageUnmaskedSecondaryImageIntensity - average fluorescence intensity of the fluorescence channel images without considering the quality mask pixelCount - average number of pixels identified as cells per image in well CellIntensitySTD - standard deviation of fluorescence channel pixel intensity of all pixels per well identified as cells by segmentation algorithm Missing values in MIDAS files are given as and empty cell or NaN License information: "UT-GPCR001 microscopy of ligand binding to M4 muscarinic receptor in live CHO-K1-hM4 cells" is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Funding: This dataser was supported by the University of Tartu ASTRA Project PER ASPERA, financed by the European Regional Development Fund, by the Enterprise Estonia Applied research programme 2021, financed by the European Regional Development Fund, by the Estonian Research Council grant (PSG230), by the COST action CA 18133 ERNEST, by the Research Training Group GRK1910 of the Deutsche Forschungsgemeinschaft (DFG), Wellcome (206194), and the Estonian Centre of Excellence in IT (EXCITE) (TK148).