title: Studying cell behavior in whole zebrafish embryos by confocal live imaging: application to hematopoietic stem cells.
abstract: 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.
authors.affiliation: Institut Curie, Centre de Recherche, PICT-IBiSA, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Renaud
authors.forename: Philippe
authors.initials: P
authors.lastname: Herbomel
authors.forename: Karima
authors.initials: K
authors.lastname: Kissa
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004625:Embryo, Nonmammalian / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D018613:Microscopy, Confocal / Q000379:methods*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics
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