Home Botany Dechorionation of Medaka Embryos and Cell Transplantation for the Generation of Chimeras
Botany JoVE (Open Access) Citable · DOI

Dechorionation of Medaka Embryos and Cell Transplantation for the Generation of Chimeras

DOI: 10.3791/2055-v
What you'll learn
  • Perform dechorionation of medaka embryos using enzymatic and mechanical techniques
  • Execute cell transplantation procedures to generate medaka chimeras
  • Mount dechorionated embryos in agarose for live imaging and microscopy
Protocol

Due to the hard chorion and soft embryos, manipulation of medaka embryos is more involved than in zebrafish. This video shows step-by-step procedures for how to manipulate medaka embryos, including dechorionation, mounting in agarose for imaging and cell transplantation for the production of chimeras. These procedures are essential to use medaka and zebrafish in a laboratory to take full advantage of their complementary features for the genetic dissection of vertebrate genome functions.

Difficulty
advanced
Total time
~2–3 hours per clutch (dechorionation + transplantation + mounting)
Model organism
Medaka (Oryzias latipes)
Biosafety
BSL-1

Steps

1
Dechorionate medaka embryos enzymatically and mechanically

Remove the hard chorion surrounding medaka embryos using enzymatic digestion followed by manual dissection under a microscope. This step prepares embryos for downstream manipulation and imaging.

▶ 01:32
2
Transplant cells between medaka embryos

Isolate donor cells and transfer them into host embryos using micromanipulation techniques to generate chimeric embryos for lineage tracing or functional studies.

▶ 04:22
3
Mount dechorionated embryos in agarose gel

Embed prepared medaka embryos in low-melting-point agarose to stabilize position for live confocal microscopy and high-resolution imaging.

▶ 06:05
4
Image chimeric embryos post-transplantation

Visualize and document transplanted cells in host embryos using fluorescence microscopy to confirm successful integration and track cell fate.

▶ 07:35
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