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.
Menétrey
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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
Dubayle
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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