How Nella's Neuron was made

A GFP labeled Rohon-Beard neuron was found in the zebrafish, embedded in soft agar.

Using a BioRad MRC600 confocal microscope, a z series was taken approximately every 30 min. The z step interval was 1.98 µm, and approximately 30-50 images were taken at each time point. An Olympus 40x water immersion objective lens was used.

The z series stacks were imported into NIH Image on a Macintosh. In the first 3-4 time points, there were a few motion artifacts (the cell was near the heart) but the rest of the image sequences were fine. The images were cropped so that the cell body was in the center of the frame. Stereo projections were done using a 7 degree tilt to left and right. The left and right images projections were imported into an RGB Photoshop file and pasted into the red and green channels respectively. The stereo images were saved as a consecutive numbered file sequence, and were made into a movie using "open image sequence" in Quicktime player pro.