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Figure 5 | BMC Developmental Biology

Figure 5

From: Multispectral fingerprinting for improved in vivo cell dynamics analysis

Figure 5

Cell Contact Analysis Using 2-Color Membrane and 1-Color Nuclear Labeling in the Embryo Resolves Cell Interactions. (A) A typical 6-8 somite embryo microinjected and electroporated with Gap 43-GFP, Gap43-YFP, and H2B-mCherry is shown. The GFP, YFP, and mCherry fluorescence signals have been pseudo-colored blue, red, and yellow, respectively, for better visual inspection. The distinct cranial NC cell migratory streams appear within the surrounding unlabeled tissue. (B-C) A higher magnification of the r4 NC cell migratory stream shows the intricate network of NC cellular processes. The arrow highlights a cell of interest at the migratory front. (D-F) The individual color channels from (C) show that not every cell is labeled with all three colors. Importantly, not all cells are labeled with both Gap43 constructs to distinguish individual cellular processes of neighboring cells. (G) A grey scale image highlights the advantage of using two different membrane labels. (H) Using semi-automated filament tracing software, NC cellular processes are rendered. However, it is difficult to distinguish between neighboring cells using both visual inspection and software. (I) Using the color information originally collected, a closer look at the same two cells (in G) reveals how the different colors highlight individual cellular processes and (J) segmentation shows the intricate network of neighboring cell contacts. All scale bars are 20 um.

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