Figure 4

Cell Tracking By Spectral Identification Resolves Cell Identity After Cell Trajectories Cross in Space and Time. (A) A typical NC cell migratory stream at with cells labeled with H2B-RFP (grayscale: the images is inverted to more clearly visualize cells). (B) A closer look at a specific group of 4 migratory neural crest cells that cross pathways shows 24 (4-factorial, 4!) different combinations of possible cell identification after the cells move apart. (C) A typical 3-color nuclear labeled embryo reveals the NC cell migratory and highlights the advantage to visually distinguish individual cell nuclei. (D-E) More specifically, tracking two dynamic migratory NC cells that cross trajectories, the color information allows both visual inspection and spectral identity to distinguish the difference of the two cells when before and after the cells cross paths. Scale bars are 10 um. Throughout a typical time-lapse imaging session with a cell trajectory crossing, the individual cells are apart (0 min), approach each other (20 min), contact each other (42 min), then start to separate (1 h 20 min and 2 h 15 min). (F-G) Line intensity profiles were drawn manually (AIM) through the center of each cell. (F' & G') The line intensity profiles shown for Cell 1 and Cell 2 show the independent spectral identity of each cell. Cell 1 has an H2B-GFP and -RFP signature, while cell 2 has H2B-RFP and -YFP signature, creating an easily distinguishable pair of cells. (H & I) After the cells cross paths, line intensity profiles show the distinct spectral identities of each cell. The scale bars are 20 um (A&C) and (D-F) 10 um.