Tissue treatment for whole mount internal lectin staining in the nematodes Caenorhabditis elegans, Panagrolaimus superbus and Acrobeloides maximus

• Published in: Histochemistry (Berlin) Vol. 101; no. 5; p. 379
• Main Authors: Borgonie, G, van Driessche, E, Link, C D, de Waele, D, Coomans, A
• Format: Journal Article
• Language: English
• Published: Germany 01.06.1994
• Subjects: Animals; Buffers; Caenorhabditis elegans - anatomy & histology; Caenorhabditis elegans - metabolism; Carbohydrate Metabolism; Freeze Etching; Glycoproteins - metabolism; Histocytochemistry; Lectins; Microscopy, Fluorescence; Microvilli - metabolism; Microvilli - ultrastructure; Nematoda - anatomy & histology; Nematoda - metabolism; Staining and Labeling; Tissue Fixation
• ISSN: 0301-5564
• DOI: 10.1007/bf00269000

Four different fixation schemes, using ten fluorescent-labelled lectins, were investigated for whole mount internal staining of three rhabditid nematodes: Caenorhabditis elegans, Panagrolaimus superbus and Acrobeloides maximus. Acetone-only fixation was found to give strong and reproducible staining, which could be prevented either by periodate treatment of the organisms or by specific inhibitory sugars of the lectins under investigation. Whereas the use of either phosphate or TRIS buffers had no effect on the staining pattern or the fluorescence intensity, the incubation time as well as the incubation temperature affected 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.