Fig. 2.

Flowchart for the concept of two-dye imaging using image stitching. In the first scan, concurrent FL target fluorescence and reflectance images were recorded, and then imported into the image stitching software to generate a two-dimensional (2-D) FL dye fluorescence hot-spot map. In the second scan, concurrent PM target fluorescence and reflectance images were recorded, and a 2-D PM dye fluorescence hot-spot map was then generated. The software then spatially registered and merged the individual FL and PM map into a combined map with FL-only, PM-only, and FL-PM mix targets.