Home›Immunology›Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining
ImmunologyJoVE (Open Access)Citable · DOI
Assessing Primary Neurogenesis in Xenopus Embryos Using Immunostaining
DOI: 10.3791/53949-v
What you'll learn
✓Mount Xenopus embryos for cryosectioning and immunofluorescent analysis
✓Prepare thin sections of embryonic neural tissue using cryosectioning
✓Identify and visualize neuronal cell populations via immunostaining
Protocol
This article presents a convenient and rapid method for visualizing different neuronal cell populations in the central nervous system of Xenopus embryos using immunofluorescent staining on sections.
Difficulty
intermediate
Total time
~4–6 hours per embryo (including mounting, sectioning, and staining)
Model organism
Xenopus laevis
Biosafety
BSL-1
Steps
1
Mount Xenopus embryos in embedding medium
Prepare and position Xenopus embryos in cryoprotectant embedding medium on cryomolds. Ensure proper orientation for cross-sectioning of the central nervous system.
▶ 00:51
2
Cryosection mounted Xenopus embryos
Use a cryostat to cut thin frozen sections of the mounted embryo. Collect sections on glass slides for subsequent immunostaining.
▶ 02:05
3
Visualize neuronal populations by immunofluorescent staining
Apply primary and secondary antibodies to embryo sections to label distinct neuronal cell populations in the neural tissue. Image stained cross-sections to assess primary neurogenesis.