Multiplexed PSF Engineering for Three-Dimensional Multicolor Particle Tracking

Three-dimensional spatiotemporal tracking of microscopic particles in multiple colors is a challenging optical imaging task. Existing approaches require trade-off between photon-efficiency, field view, mechanical complexity, spectral specificity and speed. Here, we introduce multiplexed point-spread function engineering that achieves photon efficient, 3D, multicolor particle over large view. This accomplished by first chromatically splitting the emission path microscope to different channels, each, then recombining them onto same region camera. We demonstrate our technique for simultaneously five types emitters in-vitro, as well co-localization DNA loci live yeast cells.