Two dimensional projection of a three dimensional subcellular localization plot showing the predicted relative distribution of the styryl probes in lysosomes, mitochondria and cytosol. In three dimensions, the origin of the graph would extend into the back of the page. Each axis projects towards the viewer (indicated by arrows). Each point in the plot represents an individual styryl molecule, according to its predicted, % mitochondrial, % lysosomal and % cytoplasmic (or plasma) membrane mass distribution. Each data point links to the chemical structure of each molecule and the images associated with each molecule including Hoechst, FITC, TRITC and Cy5 channels, the predicted localization and annotated staining patterns. Predicted localizations based on different input parameter values were visually compared to the staining patterns apparent in the images. Images and localization calls from 12 selected data points (numbered 1 – 12) are shown, in a range from predicted 100% mitochondrial localization to 100% cytoplasmic membrane localization, for a subset of compounds without significant lysosomal mass accumulation. Miner 3D (Miner 3D, Inc) was used to generate the figure. Additional images, chemical structures, and calculated properties for these twelve compounds are included in the (Additional file 1: Figure S1-S12).