Microscopy has been a cornerstone of discovery in the academic life sciences for more than 100 yr. This comes from a unique ability to provide extremely rich information of biological structure and dynamics. The advent of digital imaging and machine vision has brought within itself the ability to collect images more easily and critically, the ability to measure objects and intensities within images. Although many continue to use microscopy in a qualitative manner, the analytical capabilities afforded by machine vision are increasingly being applied to basic cell biology and biomedical research. Scalable quantitative imaging technology might enable scientists and engineers to determine structure, dynamics, and function of entire biological systems rather than individual molecules or pathways. This chapter will provide an overview of early efforts in the academic community to apply high content screening to the study of biological systems.