Journal
NATURE PROTOCOLS
Volume 10, Issue 12, Pages 2064-2073Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2015.130
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Funding
- Max Planck Society
- Deutsche Forschungsgemeinschaft [DFG SI-1374/3-1]
- European Research Council (ERC) [260794-ZebrafishAngio]
- Deutsche Forschungsgemeinschaft (DFG) Cells-in-Motion Cluster of Excellence, University of Munster, Germany [EXC 1003-CIM]
- [Sonderforschungsbereich (SFB) 629]
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Zebrafish possess the remarkable ability to regenerate a vast variety of tissues, even as adults. However, direct imaging of regenerative processes in adult zebrafish remains challenging because of the lack of suitable anesthesia protocols. Here we present a description of an intubation-based anesthesia procedure that we developed to enable us to image regenerating zebrafish fins and which can be used to continuously anesthetize adult zebrafish for up to 2 d. Fish are immobilized in an imaging chamber followed by oral intubation. Subsequent delivery of anesthetic-containing water is achieved via a peristaltic pump. The setup of the system will take similar to 90 min for two adult zebrafish, and it requires only a little previous experience of working with zebrafish. Our protocol will enable the imaging of regenerative processes in the fin and other tissues, and the investigation of processes that require long-term anesthesia, such as immune responses and surgical procedures.
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