TISSUE TREATMENT FOR WHOLE-MOUNT INTERNAL LECTIN STAINING IN THE NEMATODES CAENORHABDITIS-ELEGANS, PANAGROLAIMUS-SUPERBUS AND ACROBELOIDES-MAXIMUS

Four different fixation schemes, using ten fluorescent-labelled lectin s, were investigated for whole mount internal staining of three rhabdi tid nematodes: Caenorhabditis elegans, Panagrolaimus superbus and Acro beloides maximus. Acetone-only fixation was found to give strong and r eproducible staining, which could be prevented either by periodate tre atment of the organisms or by specific inhibitory sugars of the lectin s under investigation. Whereas the use of either phosphate or TRIS buf fers had no effect on the staining pattern or the fluorescence intensi ty, the incubation time as well as the in cubation temperature affecte d the staining reaction. The best results were obtained upon overnight incubation at 4 degrees C: the lectin staining could be inhibited in all cases, except for the intestinal brush border of C. elegans by the lectin of Lens culinaris.