In the last several years, multiple research publications have shown increased value of using 3D cell models compared to traditional 2D systems. Although considered by many as a more representative model of in vivo biology, setup of 3D models can be more challenging, especially when the throughput is further increased. This course will provide an overview of scaffold and scaffold free techniques and key optimizations for increased throughput through imaging technologies and automation. We will offer tips and tricks for improving cell seeding, handling, processing and use of high content imaging and analysis for 3D applications to address the need of higher throughput demands.