Light and electron microscopic detection of (3 Gal beta 1,4 GlcNAc beta 1) sequences in asparagine-linked oligosaccharides with the Datura stramonium lectin.

Abstract

The Datura stramonium lectin recognizes with high affinity the disaccharide N-acetyllactosamine (Gal beta 1,4 GlcNAc). We have developed a highly specific cytochemical affinity technique in which an ovomucoid-gold complex serves as second step reagent for the visualization of this lectin bound to reactive sequences present in tissue sections. The lectin binding sites were detected in semithin and ultrathin sections of aldehyde-fixed and low temperature Lowicryl K4M embedded tissues. For light microscopical labeling the photochemical silver reaction for signal amplification was required. The application of this technique for the detection of N-acetyllactosamine containing asparagine-linked oligosaccharides in various intracellular organelles and the plasma membrane is demonstrated.