Tissue clearing
These documents describe tissue clearing protocols relevant to our cardiovascular mouse gastruloids. Tissue clearing is used to render the tissue transparent, allowing high resolution imaging millimeters into tissue. Jana Harizanova and Sunny Dai at CFIM, University of Copenhagen, who are experts in tissue clearing techniques, found EZ Clear to be the best compromise between efficiency and accuracy for our samples when imaged with their Zeiss Light Sheet 7 microscope. Our own light sheet microscope has water dipping lenses, which gives massive spherical aberrations (image distortions) when imaging cleared samples due to refractive index mismatch. Thus, to acquire sharp 3D images using our own microscope, we instead image from different angles and combine multiview fusion with deconvolution in Huygens SVI.