Live Cell Imaging of Zebrafish Leukocytes
Zebrafish are ideally suited for the live imaging of early immune cell compartments. Macrophages that initially appear on the yolk surface prior to the onset of circulation are the first functional immune cells within the embryo, predating the emergence of the first granulocytic cells—the heterophilic neutrophils. Both cell types have been shown in zebrafish to contribute to a robust early innate immune system, capable of clearing systemic infections and participating in wound healing. Early imaging of these cells within zebrafish relied on differential interference contrast (DIC) optics because of their superficial locations in the embryo and the optical transparency of embryonic tissues. Recently, the creation of a number of transgenic reporter lines possessing fluorescently marked myelomonocytic compartments provides the potential to live image these cells during the inflammatory response, in real-time, within a whole animal context. Live imaging during the different stages of inflammation using this expanding library of reporter lines, coupled with the ability to model aspects of human disease in the zebrafish system, have the potential to provide significant insights into inflammation and diseases associated with its dysregulation.
Key wordsZebrafish Live cell imaging Neutrophils Macrophages Inflammation Phagocytosis Tg(lyz:EGFP/DsRED2) Transgenic pHrodo Escherichia coli BioParticles
The authors would like to thank Jacqui Ross and other members of the Biological Imaging Research Unit (The University of Auckland) for imaging assistance; Alhad Mahagaonkar for management of the zebrafish facility; and Annie Chien and Lisa Pullin for expert technical assistance and Makoto Kamei for imaging advice. This work was supported by a grant from the Foundation for Research Science and Technology.
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