Lorenzo Scipioni, Belén Torrado, G Tedeschi, Lila P. Halbers, Zachary R. Torrey, Erin B. Fuller, Francesco Fersini, Christoph Gohlke, Andrej Lupták, Jennifer A. Prescher, Michelle A. Digman
We report an imaging package to democratize all-in-one bioluminescence and fluorescence microscopy. The platform comprises three tools: PhasorViewer, a visualization suite to design experiments and identify optimal probe combinations; PhasorScope, an open-source, cost-effective microscopy framework to upgrade conventional microscopes; and PhasorAnalysis, a dedicated, user-friendly analysis pipeline to quantify phasor imaging data. We demonstrate the utility of the platform for multiplexed, simultaneous fluorescence and bioluminescence imaging with readily accessible optical reporters.