A simple method for the in situ detection of eosinophils

A simple and rapid method for identifying eosinophils in tissue sections is described. The assay is based on the incubation of frozen tissue sections with phenol red and the detection of cellular fluorescence. A one-to-one relationship was observed between cells exhibiting fluorescence following exposure to phenol red and eosinophils as identified by histochemical detection of eosinophil peroxidase (EPO) activity. Using the phenol red assay, eosinophils were detected in various eosinophil-infiltrated tissues, including uteri of rats from day 1 of pregnancy and uteri of prepubertal estrogen-treated rats. Intensity of the fluorescing eosinophils was dependent upon phenol red concentration and duration of incubation. The phenol red method of eosinophil identification was disrupted by co-incubation with resorcinol, an EPO inhibitor, or catalase, a hydrogen peroxide scavenger. The merits of this assay are its simplicity and compatibility with other procedures, such as histochemistry, immunocytochemistry, and in situ ligand-receptor binding.