An inexpensive programmable illumination microscope with active feedback

We have developed a programmable illumination system capable of tracking and illuminating numerous objects simultaneously using only low-cost and reused optical components. The active feedback control software allows for a closed-loop system that tracks and perturbs objects of interest automatically. Our system uses a static stage where the objects of interest are tracked computationally as they move across the field of view allowing for a large number of simultaneous experiments. An algorithmically determined illumination pattern can be applied anywhere in the field of view with simultaneous imaging and perturbation using different colors of light to enable spatially and temporally structured illumination. Our system consists of a consumer projector, camera, 35-mm camera lens, and a small number of other optical and scaffolding components. The entire apparatus can be assembled for under $4,000. Supplemental matlab code is available to assist in the setup of the active feedback software.