A one-step dual-labeling method for antigen detection in mast cells.
This paper describes a one-step light microscopy method for demonstrating the antigen contents of unequivocally identified mast cells. It is based on the differential metachromatic properties of proteoglycans, mostly heparin and chondroitin sulfate, and 1-naphthol in the presence of toluidine blue in an acidic medium. Proteoglycans occur in all mast cells and 1-naphthol is used to demonstrate the peroxidase activity of the sections treated by the horseradish peroxidase-labeled avidin-biotin complex method for antigen detection. Granules containing proteoglycans present the classical metachromatic reaction by appearing purplish-red, while granules containing antigen appear a brilliant green. When both types of granules are distinct inside the cell, single- and double-stained cells can be accurately separated and counted. We hope that this new procedure will contribute to a further identification of mast cell mediator contents and to a better understanding of the physiology of this cellular population.
CNRS UMR 8119 Neurophysique et physiologie du système moteur, UFR Biomédicale, Université René Descartes, 45 rue des Saints Pères, 75270, Paris Cedex 06, France. daniel.menetrey@biomedicale.univ-paris5.fr
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D000818:Animals; D000941:Antigens / Q000032:analysis*; D002809:Chondroitin Sulfates / Q000032:analysis; D004396:Coloring Agents; D005404:Fixatives; D005557:Formaldehyde; D006493:Heparin / Q000032:analysis; D006632:Histamine / Q000032:analysis; D007150:Immunohistochemistry / Q000379:methods*; D008407:Mast Cells / Q000737:chemistry*; D009284:Naphthols; D051381:Rats; D012701:Serotonin / Q000032:analysis; D014048:Tolonium Chloride
D000941:Antigens; D004396:Coloring Agents; D005404:Fixatives; D009284:Naphthols; D014048:Tolonium Chloride; D005557:Formaldehyde; C029350:1-naphthol; D012701:Serotonin; D006632:Histamine; D006493:Heparin; D002809:Chondroitin Sulfates