Transgenic gfap:eGFP embryos were injected at the one cell stage with mRNA encoding nuclear localized RFP. Z-stacks were imaged using high speed resonant scanning with a confocal microscope positioned dorsally over the neural tube starting at 23hours post fertilization. Every five minutes a new stack was imaged for ~3h at a constant temperature of 28.5° C.

One cell (asterisk positioned on red nucleus) that was GFP positive underwent one cell division during a period that was less then 1h and 40 minutes (two asterisks at t31). Timelapse movie is seen at three frames per minute and positioned constantly on one plane of the Z-stack acquired. Movie generated with Volocity.

Transgenic gfap:eGFP embryos were injected at the one cell stage with mRNA encoding nuclear localized RFP. Z-stacks were imaged using high speed resonant scanning with a confocal microscope positioned laterally over the neural tube starting at 23hours post fertilization. Every three minutes a new stack was imaged for ~3h at a constant temperature of 28.5° C.

One cell (asterisk positioned on red nucleus) that was negative for GFP underwent one cell division during the 3h period (two asterisks at t37). Timelapse movie is seen at three frames per minute and positioned for a lateral view of the neural tube positioned constantly on one plane of the Z-stack acquired. Movie generated with Volocity.

This is a laser Scanning Confocal Z-Stack projected to rotate around the Y axis. This image is a frontal view of the zebrafish forebrain at 30h. Antibody labeling for Acetylated Tubulin (red) labels all axons in the forebrain. The anterior (top) and Postoptic commissures (POC, bottom) can be seen. Cells from the animal cap of a gastrula staged gfap:gfp transgenic embryo were transplanted into a non-GFP transgenic line.

The green cells are the surviving transplanted cells that took root in the forebrain. Appropriately these cells generated a radial glial morphology. This is the first time we have actually been able to see clear morphology of the entire cell.

This movie was made using the Volocity 4D rendering software. This software allows us to “enter” into the labeling and see cellular morphology and cell to cell interactions from all perspectives.

This is a laser Scanning Confocal Z-Stack projected to rotate around the Y axis. This image is a frontal view of the zebrafish forebrain at 40h. Antibody labeling for Acetylated Tubulin (red) labels all axons in the forebrain. The anterior (top) and Postoptic commissures (POC, bottom), as well as the optic chiasm (just over the POC) can be seen. gata2:gfp positive POC neuronal precursor cells were transplanted from an 18h old embryo to a non-GFP transgenic line.

The green cells are the surviving transplanted cells that took root in the forebrain and appropriate grew axons along the POC (green lines going from left to right across the midline.

This represents flattened laser scanning Confocal Z-Stacks taken over a 2h period. Transplanted cells are red and green nuclei are cells expressing GFP driven by the gfap promoter. This movie is focused on the forebrain from a frontal view at 30h.

This is the same timelapse series as shown above, but rotated to provide a different perspective.

This movie was made using the Volocity 4D rendering software. This is the same timelapse series as shown two movies above. As this timelapse runs the 3D image will rotate about the X axis.

This movie was made using the Volocity 4D rendering software. This is the same timelapse series as shown three movies above. As this timelapse runs the 3D image will zoom into a small cluster of cells and then run.