ZFIN ID: ZDB-PUB-111122-15
Studying cell behavior in whole zebrafish embryos by confocal live imaging: application to hematopoietic stem cells
Renaud, O., Herbomel, P., and Kissa, K.
Date: 2011
Source: Nature Protocols   6(12): 1897-1904 (Journal)
Registered Authors: Herbomel, Philippe, Kissa-Marin, Karima
Keywords: cell culture, model organisms, tissue culture
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Embryo, Nonmammalian/cytology*
  • Hematopoietic Stem Cells/cytology*
  • Microscopy, Confocal/methods*
  • Zebrafish/embryology*
  • Zebrafish/genetics
PubMed: 22082984 Full text @ Nat. Protoc.
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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.
ADDITIONAL INFORMATION