Smz1000 stereoscope

Zebrafish embryos were anaesthetized with 0.006% (v/v) Tricaine, placed dorsally on separate cavities of a glass slide (Marienfeld Superior, 10622434), mounted on 1,2% low-melting agarose and a drop of E3 embryonic water was added on top of semi-solidified mounting medium for maintenance of humidity. For in vivo imaging, fluorescent and brightfield videos of 10sec were recorded by microscope inverted Leica DMIRE2 with a mounted Hamamatsu ORCA-Flash4.0 camera. Confocal imaging was performed using a Leica TCS SP5 II on a DM 600 CFS Upright Microscope. The images were captured with the LAS AF software, analyzed in ImageJ Software and presented as maximum projection of a set of z-stacks for each stained tissue section. Adult zebrafish were euthanized in 0.016% tricaine containing 0.1M potassium chloride to arrest the heart chambers in diastole 34 . Images of whole hearts were captured using DFK2BUC03 camera from The Imaging Source mounted on SMZ1000 stereoscope. Then, adult hearts were fixed in 4% paraformaldehyde at 4℃ overnight, washed three times with PBS, dehydrated in EthOH, and embedded in paraffin. Paraffin sections of 5μm thickness were performed using Leica RM2265 microtome. Haematoxylin and Eosin staining according to standard laboratory protocols. Images of stained sections were capture with Leica DFC500 camera mounted on Leica DMLS2 microscope.