✓Perform zebrafish gastrula stage cell transplantation using micromanipulation techniques
✓Prepare and label donor cells with fluorescent dextran for tracking
✓Image and visualize transplanted cells in developing zebrafish embryos
✓Generate tissue-specific chimeras to study developmental processes in forebrain
Protocol
Zebrafish cell transplantation enables the combination of genetics and embryology to generate tissue specific chimeras. This video demonstrates gastrula staged cell transplantations that have allowed our lab to investigate the roles of astroglial populations and specific guidance cues during commissure formation in the forebrain.
Difficulty
advanced
Total time
~4–6 hours per experimental batch (including embryo preparation, transplantation, and live imaging)
Model organism
Zebrafish (Danio rerio) gastrula stage embryos
Biosafety
BSL-1
Steps
1
Prepare microinjection plate and glass needles
Assemble and calibrate the microinjection apparatus, including needle fabrication and plate setup for cell aspiration and transplantation.
▶ 01:22
2
Microinject fluorescent dextran into donor cells
Load Alexa Fluor 594 dextran into micropipette and inject into target donor cells to enable fluorescent visualization post-transplantation.
▶ 06:03
3
Set up embryo mounting and transplant apparatus
Position gastrula-stage zebrafish embryos on agarose plates and arrange micromanipulation equipment for cell transplantation under dissecting microscope.
▶ 07:03
4
Perform gastrula stage cell transplants
Aspirate labeled donor cells with micropipette and transplant into defined tissue regions of host gastrula embryos using micromanipulation.
▶ 10:01
5
Visualize and image transplanted embryos
Recover transplanted embryos in culture medium and perform confocal or fluorescence imaging to document donor cell integration and migration patterns.
▶ 13:26
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