Ruthenium red and alcian blue effects on outer structures of methanotrophic bacteria

Abstract

Methane, a contributor to the “greenhouse effect”, is oxidized in the natural environment by methanotrophic bacteria. As part of a comprehensive research effort, we have been examining the ultrastructure of methanotrophs. These microorganisms have complex outer cell wall structures similar to those frequently found in other chemol itho- trophic bacteria. (1,2)In our work, we have focused on the “type” strains of Methylomonas albus BG8 and Methylosinus trichosporium OB3b. Between Spurr and LR White embedding resins, we found a difference 1n the preservation of an outer cup layer of BG8 external to the peripheral membranes. Cells from the same sample embedded in Spurr consistently lacked this feature (FIG. 1). This effect was overcome by an en bloc ruthenium red (RR) protocol that resulted in successful retention of the cup layer in Spurr resin (FIG. 2). For OB3b cells, the en bloc RR protocol resulted in an exterior bead feature distinguishable in thin section (FIG. 4) that previously was seen only by SEM.