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Ex Vivo Oculomotor Slice Culture from Embryonic GFP-Expressing Mice for Time-Lapse Imaging of Oculomotor Nerve Outgrowth
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1Department of Ophthalmology,Boston Children’s Hospital, 2Department of Ophthalmology,Harvard Medical School, 3F.M. Kirby Neurobiology Center,Boston Children’s Hospital, 4Department of Neurology,Boston Children’s Hospital, 5Department of Neurology,Harvard Medical School, 6Howard Hughes Medical Institute
Chapters
- 00:04Title
- 00:51Pregnancy Confirmation and E10.5 Embryo Harvest and Slice Preparation
- 04:15Results: Representative Ex Vivo Oculomotor Slice Culture Development and Pitfalls
- 05:30Conclusion
An ex vivo slice assay allows oculomotor nerve outgrowth to be imaged in real time. Slices are generated by embedding E10.5 IslMN:GFP embryos in agarose, slicing on a vibratome, and growing in a stage-top incubator. The role of axon guidance pathways is assessed by adding inhibitors to the culture media.
