NDLI: In ovo live imaging of avian embryos.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Kulesa, Paul M. Bailey, Caleb M. Cooper, Cameron Fraser, Scott E.
Description	Country affiliation: United States Author Affiliation: Kulesa PM ( Stowers Institute for Medical Research, Kansas City, MO 64110, USA. pmk@stowers.org)
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 (approximately 36 h). This technique has been adapted to study somitogenesis.
File Format	HTM / HTML
ISSN	19403402
e-ISSN	15596095
DOI	10.1101/pdb.prot5446
Journal	Cold Spring Harbor Protocols
Issue Number	6
Volume Number	2010
Language	English
Publisher	Cold Spring Harbor Laboratory Press
Publisher Date	2010-06-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Clinical Laboratory Techniques Embryology Embryo, Nonmammalian Imaging, Three-dimensional Ovum Animals Calibration Chick Embryo Cytology Embryonic Development Hot Temperature Membranes, Artificial Growth & Development Polytetrafluoroethylene Quail Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Biochemistry, Genetics and Molecular Biology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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