Figure 4.

Component-wise comparison of spectral unmixing using sub-selected excitation wavelength images. From left to right, the columns show false-colored, unmixed images for center wavelength spacing every 5 nm, every 20 nm, every 40 nm, and specially chosen to correspond to “peak wavelengths” as described in Figure 3. From top to bottom, components include autofluorescence, Fura-2, Fura Red, MitoTracker Green, NucBlue, and TRITC WGA. The penultimate row contains the composite images generated from merging the images above them. The bottom row shows the visualized RMS error.