Use of a fluorescent brightener to demonstrate cellulose in the cellular slime molds

Abstract

The presence and location of cellulose in different stages of the life cycles of the cellular slime molds can be demonstrated by use of the disodium salt of 4,4'-bis(4-anilino-6-bis (2-hydroxyethyl)-amino-s-triazin-2-ylamino)-2,2' -stilbene disulfonic acid, a fluorescent brightener. It may be used successfully as a direct stain at a concentration of 0.1% in half-normal saline at pH 6; and it may be incorporated into growth media as a vital stain at a concentration of 0.0025% with no inhibitory effect at any developmental stage. Vegetative myxamoebae contain no cellulose and show no fluorescence in the presence of this brightener when viewed with ultraviolet light. In later stages of the life cycle, the time and sites of cellulose formation can be demonstrated with the brightener because of its fluorescence. e.g., in the slime covering of the pseudoplasmodia, in the sorophore sheath, in the walls of stalk cells and spores, in the walls of microcysts, and in the walls and sheath material of macrocysts. The brightener appears to be a very sensitive indicator for cellulose, and it has certain advantages over other cellulose stains, since the staining reaction (fluorescence) is very intense, long-lasting, and not obscured by unstained cellulose-free myxamoebae if such are present.