Spot rt3 camera

Fin clipped larvae were fixed in 4% PFA at 4 °C overnight and kept in 70% ethanol. At least five embryos or larvae per genotype group were embedded in 1% agarose in 1X TAE buffer. A mould, specifically designed to align zebrafish larvae, was used to produce agarose blocks with identical distributed wells of the same depth. Agarose blocks were gradually dehydrated in an enclosed automated tissue processor (Shandon Excelsior ES, Thermo Scientific) and subsequently embedded in paraffin. The heads of paraffin-embedded larvae were sectioned on a HM 325 manual rotary microtome (Thermo Fisher Scientific) at a thickness of 5 μm. The specimens were stained with hematoxylin and eosin (H&E stain) using Varistain™ Gemini ES Automated Slide Stainer (Thermo Fisher Scientific) according to laboratory protocols. The resulting sections were imaged at ×20 magnification in a SPOT 5.1 software (SPOT Imaging) by a SPOT-RT3 camera mounted on a Leica microscope. Brightness of the images was adjusted for the white background.

VHC- and KA-injected larvae were fixed in 4% paraformaldehyde (PFA) at 4°C overnight and kept in 70% ethanol. At least four larvae per genotype group were embedded in 1% agarose in 1x TAE buffer. A specifically designed mold was used to produce agarose blocks with identically distributed wells of the same depth. Agarose blocks were gradually dehydrated in an enclosed automated tissue processor (Shandon Excelsior ES, Thermo Scientific) and subsequently embedded in paraffin. Heads of paraffin-embedded larvae were sectioned on a HM 325 manual rotary microtome (Thermo Fisher Scientific) at 5 μm thickness. Specimens were stained with haematoxylin and eosin (H&E) using Varistain™ Gemini ES Automated Slide Stainer (Thermo Fisher Scientific). Resulting sections were imaged at 20x and 40x magnification in SPOT 5.1 software (SPOT Imaging) by a SPOT-RT3 camera mounted on a Leica microscope. Brightness of the images was adjusted for the white background.