The use of vibratome sections for the ruthenium tetroxide protocol: a key for optimal visualization of epidermal lipid bilayers of the entire human stratum corneum in transmission electron microscopy.

In this paper we describe a preparative procedure which allows maximal visualization of all lipid bilayers in the human stratum corneum (SC). We used 50-microns-thick vibratome sections of paraformaldehyde/glutaraldehyde-fixed human skin. The sections were post-fixed in 1% osmium tetroxide and 0.5% ruthenium tetroxide. The vibratome sections were dehydrated only in 70% ethanol in order to prevent dissolution of the lipids. Lipid bilayers, including the alternating electron-dense and electron-lucent lamellae, were visible between all cell layers of the SC. In addition, this preparative procedure also appeared to be excellent for the ultrastructural preservation of lamellar bodies.