Histochemical heterogeneity of mast cells in rat dermis

The dermal mast cells of Wistar rats were studied following fixation in either 10% phosphate buffered formalin or Carnoy's solution and staining with either toluidine blue or alcian blue:safranin O. Granules of mast cells appeared heterogeneous following fixation with formalin and staining with alcian blue:safranin O, but not when stained with toluidine blue. The number of mast cells observed in skin fixed in Carnoy's solution was greater than the number observed in equivalent samples of skin fixed in formalin (p < 0.01) when both samples were stained with toluidine blue. In formalin fixed skin stained with alcian blue:safranin O, there were three populations of mast cells designated as "blue," "red" or "mixed." "Blue" mast cells, containing only alcian blue stained granules, "red" mast cells, containing only safranin O stained granules, and "mixed" mast cells, containing both alcian blue and safranin O stained granules accounted for 77.6 +/- 3.0, 6.6 +/- 2.5 and 15.8 +/- 2.5% of the total mast cell population, respectively. In skin specimens fixed in Carnoy's solution and stained with alcian blue:safranin O the mast cells contained only blue granules (alcian blue positive). The number of mast cells observed in Carnoy's fixed skin sections was less than the number seen in formalin fixed skin sections when both were stained with alcian blue/safranin O. This indicates that there is a group of mast cells which do not stain with safranin O after fixation with Carnoy's solution.