This is a Shared Instrument Grant proposal for the acquisition of a high content imaging system, the Perkin Elmer Opera Phenix Plus to replace three discontinued IN Cell high content systems which will no longer be supported as of 2025. The current three IN Cell instruments reside in three important Core facilities at the Mission Bay Campus of the University of California, San Francisco (UCSF). High content imaging is a key service for many investigators at UCSF who use the core services of the Small Molecule Discovery Center (SMDC), the Center of Cellular Construction CCC) and the Environmental Research and Translation for Health (EaRTH). Many of our investigators have research projects that screen for morphological changes in a high- throughput manner for varying conditions (e.g. pH, gene knockouts, environmental chemicals, small molecule drugs) in a wide variety of biological samples (e.g. mammalian tissues and organoids, Stentor, budding yeast, nematodes, zebrafish, neurons and phase separation droplets) using high content image-based screening. The Opera Phenix Plus system can act as a replacement for all three IN Cell instruments due to its simultaneous 4-camera system that reduces the acquisition time such that it is at least four times faster than the top of the line IN Cell 6000 and 6500 that reside currently in our Cores. With the faster throughput, the new instrument will be able to accommodate all the users from the three different Cores. In addition, the faster acquisition will allow 3D time-lapse imaging that many of our investigators attempted on our IN Cells but were not satisfied with time it took to complete a 96-well plate, typically over an hour for a typical sample compared to twelve minutes with the Opera Phenix Plus. The new instrument also is equipped with water immersion lenses for 20, 40 and 63x magnification, which due to better refractive index matching, gives higher resolution images than could be obtained in the IN Cell instruments which relied on air objectives. The Opera Phenix Plus is also equipped with lasers for multi-wavelength imaging, environmental control, automation and easy-to-use imaging processing software that are needed for our investigators to successfully conduct their research. The innovative research programs of our users span a wide range of basic, translational, and clinical research areas as well as address many health-related areas in the realms of cancer, developmental, reproductive and neurological disorders. All 17 Major and Minor users have NIH grants that support their high content imaging. The Opera Phenix Plus will be essential to continue and further develop our biomedical research for many years to come.