Studying cell behavior in whole zebrafish embryos by confocal live imaging: application to hematopoietic stem cells
- Renaud, O., Herbomel, P., and Kissa, K.
- Nature Protocols 6(12): 1897-1904 (Journal)
- Registered Authors
- Herbomel, Philippe, Kissa-Marin, Karima
- cell culture, model organisms, tissue culture
- MeSH Terms
- Animals, Genetically Modified
- Embryo, Nonmammalian/cytology*
- Hematopoietic Stem Cells/cytology*
- Microscopy, Confocal/methods*
- 22082984 Full text @ Nat. Protoc.
Renaud, O., Herbomel, P., and Kissa, K. (2011) Studying cell behavior in whole zebrafish embryos by confocal live imaging: application to hematopoietic stem cells. Nature Protocols. 6(12):1897-1904.
Confocal live imaging is a key tool for studying cell behavior in the whole zebrafish embryo. Here we provide a detailed protocol that is adaptable for imaging any progenitor cell behavior in live zebrafish embryos. As an example, we imaged the emergence of the first hematopoietic stem cells from the aorta. We discuss the importance of selecting the appropriate zebrafish transgenic line as well as methods for immobilization of embryos to be imaged. In addition, we highlight the confocal microscopy acquisition parameters required for stem cell imaging and the software tools we used to analyze 4D movies. The whole protocol takes 2 h 15 min and allows confocal live imaging from a few hours to several days.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes