Tcs sp8 hyd

Immediately after microinjection, anesthetized larvae were mounted in custom-made 2% agarose mold and covered with 1% low melting point agarose. After solidifying, the mold was covered with egg water plus tricaine. The sample was then taken to a Leica TCS SP8 HyD inverted confocal microscope for live imaging. We used a 40× oil immersion objective (NA 1.3). Time-lapse videos, from a total of six zebrafish larvae, were processed and analyzed using Fiji open-source software, version 2.9.0/1.53t [34 (link)].

Images were captured with a Zeiss LSM710 META BIG, Zeiss LSM 710 FCS or Leica TCS SP8 HyD confocal microscopes with the 10×, 20× or 40× oil objectives. Images were analyzed using the Bitplane IMARIS suite and Image J. Imaging was performed in the Australian Cancer Research Foundation (ACRF)'s Dynamic Imaging Facility at the Institute for Molecular Bioscience (University of Queensland) and the Sydney Cytometry facilities at Centenary Institute (University of Sydney). All graphs and statistical tests were performed with Graphpad Prism 9 and illustrated with Adobe Illustrator CS6. Morphological, cell count, and proliferation experiments were imaged using a Leica SP8 inverted microscope at Monash Microimaging platform.