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In Ovo Live Imaging of Avian Embryos

Kulesa, P. M. and Bailey, C. M. and Cooper, C. and Fraser, S. E. (2010) In Ovo Live Imaging of Avian Embryos. Cold Spring Harbor Protocols, 2010 (6). Art. No. 5446. ISSN 1940-3402. PMCID PMC3491899. https://resolver.caltech.edu/CaltechAUTHORS:20160106-092837460

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Abstract

Vertebrate development is best studied in an intact embryo model, but a robust interface between time-lapse microscopy and in vivo embryo health and maintenance can be difficult to achieve in model systems that rely on external factors for life support. This protocol presents a system for in ovo culture and time-lapse imaging of fluorescently labeled cells within living avian embryos, using a Teflon membrane that is oxygen-permeable and liquid-impermeable. The protocol describes the Teflon membrane assembly (the assembly size can be changed to fit smaller eggs, such as those of the quail), its interface with the egg window, and the use of an upright microscope and heated chamber. The use of the system is demonstrated in chick embryos by following individual fluorescently labeled neural crest cells, a multipotent stem cell-like population that differentiates into a wide range of derivatives and travels extensively throughout the embryo. By combining in ovo culture with confocal or two-photon four-dimensional time-lapse imaging, embryo health can be maintained for up to 5 d, and neural crest cell behaviors can be visualized for long periods of time (~36 h). This technique has been adapted to study somitogenesis.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1101/pdb.prot5446DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3491899/PubMed CentralArticle
ORCID:
AuthorORCID
Fraser, S. E.0000-0002-5377-0223
Additional Information:© 2010 Cold Spring Harbor Laboratory Press. The Authors acknowledge that six months after the full-issue publication date, the Article will be distributed under a Creative Commons CC-BY-NC License (Attribution-NonCommercial 4.0 International License, http://creativecommons.org/licenses/by-nc/4.0/). We thank Dan Kiehart (Duke University) for the initial ideas of using Teflon membranes in embryo culture for intravital imaging. P.M.K. thanks the National Institutes of Health (NIH; 1R01HD057922) and the Stowers Institute for Medical Research for funding. S.E.F. thanks NIH (P50HG004071) and the Beckman Institute for funding.
Funders:
Funding AgencyGrant Number
NIH1R01HD057922
Stowers Institute for Medical ResearchUNSPECIFIED
NIHP50HG004071
Caltech Beckman InstituteUNSPECIFIED
Issue or Number:6
PubMed Central ID:PMC3491899
Record Number:CaltechAUTHORS:20160106-092837460
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160106-092837460
Official Citation:Kulesa PM, Bailey CM, Cooper C, Fraser SE. In Ovo Live Imaging of Avian Embryos. Cold Spring Harbor protocols. 2010; 2010(6):pdb.prot5446. doi:10.1101/pdb.prot5446.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:63411
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:06 Jan 2016 17:45
Last Modified:03 Oct 2019 09:28

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