Video Gallery

The Barresi lab principally generates data using fluorescent microscopy. The images, 3D projections and timelapse movies presented on this page were either collected using structural illumination from an epifluorescent microscope or using a laser scanning confocal microscope.

Currently, the movies provided on this page are largely focused on characterizing glial cells of the forebrain or cell division occurring specifically in astroglia of the spinal cord. All images and movies are the property of the Barresi Lab. Usage requires permission from Dr. Michael Barresi.

1. Resonant Laser Scanning Confocal Timelapse of POC formation

Frontal view of the ventral forebrain of live gap43:Gfp transgenic embryos from 23h to 25h of development. Z-stacks were taken every 4minutes and then compiled into this timelapse movie. The first growth cones detected are actually derived from neurons pioneering the AC, however these quick retract. Appropriately timed, POC pioneering growth cones located lower in this movie are seen pathfinding toward the midline from both sides of the brain. They converge at the midline and then continue to grow along each other in opposite directions. After this occurs the pioneering AC axons then begin to approach the midline as well. The migrating cell that enters the field from the bottom is unknown at this time.

2. Timelapse imaging of astroglial cell division in the zebrafish neural tube

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.

3. Timelapse imaging of cell division in a non-astroglial cell type in the zebrafish neural tube

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.

4. Laser Scanning Confocal Z-Stack projected to rotate around the Y axis: frontal view of the zebrafish forebrain at 30h.

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.

5. Scripted movie of the projection of gfap:gfp gastrula stage transplant double labeled for Axons

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.

6. Projection of gata2:gfp late stage transplant double labeled for Axons

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.

7. 4D reconstruction of gfap:gfp transplanted cells in a live gfap:nuclear localized gfp transgenic embryo over time

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.

8. A magnified region of the 4D reconstruction of gfap:gfp transplanted cells in a live gfap:nuclear localized gfp transgenic embryo over time

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.