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.
van der Meulen
J
J
Laboratory for Electron Microscopy, Leiden University, The Netherlands.
van den Bergh
B A
BA
Mulder
A A
AA
Mommaas
A M
AM
Bouwstra
J A
JA
Koerten
H K
HK
D004817:Epidermis / Q000648:ultrastructure*; D006801:Humans; D008051:Lipid Bilayers; D008854:Microscopy, Electron; D017975:Ruthenium Compounds
D008051:Lipid Bilayers; D017975:Ruthenium Compounds; C027646:ruthenium tetraoxide