The "UT-GPCR001 microscopy of ligand binding to M4 muscarinic receptor in live CHO-K1-hM4 cells" dataset contains the raw microscopy images of the experiments along with images processed using the random forest algorithm and U-Net3 based deep convolutional neural networks for cell segmentation from bright-field images. The dataset contains experiments with a fluorescence ligand UR-CG072 and multiple M4 muscarinic receptor ligands. The dataset contains binding kinetic, fluorescence ligand saturattion and competition type experiments.