Immunocytochemistry for NGF in eosinophil sections.

Freshly isolated eosinophils were collected and mixed with 1% agar and embedded in paraffin. Five-micrometer sections were blocked with normal rabbit serum and incubated with goat antihuman NGF antibody, followed by additional blocking with chromotrope 2R. Bound antibody was then visualized by affinity-purified FITC-conjugated rabbit antigoat IgG. The fluorescence images (upper panel) were captured with a confocal laser-scanning microscope. Transmission images (lower panel) were also captured from the same field to visualize the morphology of the cells. Note that the fluorescence image is captured from an approximately 1-μm optical section and that the transmission image is from a whole 5-μm section. Original magnification × 600.