*3.6. Tissue Eosinophil Counts*

At 48 h after being challenged, mice were sacrificed, and the ears were removed and fixed with 4% paraformaldehyde in a 0.1 M Phosphate buffer, pH 7.2. Then, the ears were embedded in paraffin, and sections of ears were stained with hematoxylin/eosin solution. The number of eosinophils infiltrated into the dermis was examined at a magnification of 400×. At least 10 fields were examined for each ear lobe. The number of eosinophils was counted and expressed as the number of cells in a field 

(200 ΐm × 200 ΐm). 

## *3.7. Percentage of Eosinophils in Peripheral Blood*

At 48 h after the antigen challenge, mice were bled from the retro-orbital plexus. The percentage of eosinophils among the white blood cells was determined by making a smear on a slide glass for each mouse. Slide glasses were stained with Giemsa solution, and the percentage of eosinophils among the white blood cells was estimated by counting eosinophils and all white blood cells in the field until at least 200 white blood cells were counted in each sample. At a minimum, 10 slides were counted and the mean of the counts were taken for each group. 
