Histamine immunohistochemistry: a new and highly sensitive method for studying cutaneous mast cells.
Mast cells have characteristic granulae containing various glucoseaminoglycans, proteases and amines (predominantly histamine). The conventional histological methods for studying mast cells are based upon acidic ortho- and metachromatic routine stains of the glucoseaminoglycans. However, the success of these procedures is dependent upon both the fixatives and the tissues used. In this study, we wanted to find out whether an immunohistochemical procedure could overcome some of these difficulties. Normal human skin was fixed in five different types of fixative and processed for indirect immunofluorescence, using an antiserum to histamine. Only one, 4% carbodiimide in 0.1 M phosphate buffer (pH 7.4), resulted in immunostaining. The quality of the staining was good, with a high signal-to-noise ratio, and was located on the mast cells. The method made it possible to visualize small structures such as a single secreted granula, the thin cytoplasmatic extension of some cells, and a previously undescribed dendritic morphology of some of the mast cells. We therefore recommend this procedure for cellular studies of mast cells when accuracy is needed.
Johansson
O
O
Department of Histology and Neurobiology, Karolinska Institutet, Stockholm, Sweden.
Virtanen
M
M
Hilliges
M
M
Yang
Q
Q
D000328:Adult; D002234:Carbodiimides; D005455:Fluorescent Antibody Technique; D006632:Histamine / Q000032:analysis*; D006801:Humans; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D008407:Mast Cells / Q000737:chemistry / Q000166:cytology*; D012867:Skin / Q000737:chemistry / Q000166:cytology*
D002234:Carbodiimides; D007202:Indicators and Reagents; D006632:Histamine