Organoid, an in vitro 3D culture, has extremely high similarity with its source organ or tissue, which creates a model in vitro that simulates the in vivo environment. Organoids have been extensively studied in cell biology, precision medicine, drug toxicity…which have been proven to have high research value. Periodic observation of organoids in microscopic images is essential for organoid research but it is difficult and time-consuming to perform manual screens for organoids. It’s in this way, to help research, we would like to have the opportunity to present our workshop at Mifobio First, to show a potential application in cancer research using organoid technology to facilitate drug testing. Indeed, if organoids still fail to model immune system accurately, we can demonstrate the use of co-cultures between organoids and lymphocytes after different immune-treatments and the read-out associated: 1) Lymphocytes recruitment (migration of lymphocytes inside the tumor) 2) Lymphocytes engagement (analyze of granzyme or perforin marker) 3) Positive response (Decrease of organoids size or proliferation status by KI67 marker) To do this, we’ll show the implementation of the workflow to prepare the samples (description of the material & method) and in the second part to detect, acquire and analyze automatically organoids in high-throughput assays. Indeed, the automated acquisition and detection should be realized thanks to a Spinning Disk microscope from Olympus using the associated CellSens imaging software. For the images analysis, we ‘ll perform a spatial 3D analysis at cellular and subcellular level using different machine learning methods from Imaris software.