This video describes a microinjection method for breast cancer cell delivery into the perivitelline space of transgenic zebrafish embryos. After incubation, we visualize the embryo under a fluorescent microscope to observe cancer cell invasion, dissemination, and metastasis.
Protocol
1. Image and Analyze the Metastatic Process Collect several anesthetized embryos with a wide-tip Pasteur pipette and transfer them to the glass bottom of a polystyrene dish. Remove excess water and keep a limited amount of egg water. Manipulate the embryo into position with a hair loop tool and place a cover on top of the glass. Use an inverted confocal microscope in combination with water-immersion or long-distance dry objectives. Position the embryo such that the region …
Offenlegungen
The authors have nothing to disclose.
Materials
Wide-tip Pasteur pipette (0.5-20 µL)
Eppendorf
F276456I
Confocal microscope
Leica
SP5 STED
Stereo microscope
Leica
MZ16FA
Tg (fli:EGFP) zebrafish strain
Kindly provided by Dr. Ewa SnaarJagalska (Institute of Biology, Leiden University, Leiden, The Netherlands)
zPDX-Analysis of Invasiveness: Investigating Invasive Behavior of Metastatic Cancer Cells in Zebrafish Embryo Xenografts. J. Vis. Exp. (Pending Publication), e20219, doi: (2023).