Heavy metal--a contrasting substance for micro-tomographical visualization of scorpion book lungs

Remarkable results obtained while searching for suitable contrasting agents for tomographical imaging are presented. Book lungs, the respiratory organs of scorpions (Scorpiones, Arachnida), served as challenging objects in terms of fragility and preparatory requirements, as well in terms of size differences between the entire organ and its functional subunits (the lamellae). The problem of tissue contrast was investigated using various metals, but most successfully addressed using osmium. Gaseous osmium tetroxide exposure with subsequent ethanol treatment yielded the best results. A constant and thorough staining of the book lungs was achieved, easing book-lung visualization using 3D-reconstructions from microCT scans.