JoVE Encyclopedia of Experiments
Neuroscience
0 views • 4:49 min • June 17th, 2025
Begin with a live, dechorionated transgenic zebrafish embryo embedded in low-melting-point agarose and placed in a chamber filled with embryo media.
The embryo expresses enhanced green fluorescent protein (EGFP)-labeled neural crest cells.
Position the chamber on the microscope stage. Using bright-field illumination, adjust the objective to focus and center the embryo in the field of view.
Switch to the water-immersion objective and refocus on the embryo.
Adjust the imaging parameters to acquire Z-stack images from the lateral to the medial edge, capturing the width of the eye.
Turn off bright-field illumination, cover the stage, and initiate time-lapse imaging.
Refill the chamber with media as needed during imaging.
Under laser illumination, the neural crest cells fluoresce.
Visualize the migration of these cells from the neural tube into the craniofacial regions, around the developing eye to populate the periocular mesenchyme and frontonasal process, and ventrally to form the pharyngeal arches.
After placing the entire setup onto the stage of the microscope, according to the text protocol, use the five times objective to locate the embryo. Then manually raise the stage to the highest pos
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