Androgen withdrawal increases medullary and cortical epithelial cell area 3 weeks after castration. Thymi were removed 3 weeks after castration or sham castration from 6 to 7 mice per group. Immunohistochemistry analysis of frozen sections using laser scanning cytometry was performed on 5 levels for each thymus, 2 or more slides per level, a minimum of 10 sections per mouse, 6 mice per group, were analyzed. Thus, data represent average area of 12 to 21 slides per level. (A) Representative laser scanning cytometry pseudoplot of K8⁺ cortical area (Texas Red) and K5⁺ medullary area (FITC) for sham-castrated thymus (left) and castrated thymus (right). To the right of the pseudoimage, the corresponding dot-plot for FITC-K5 versus Texas Red-K8 fluorescence is shown. The table below summarizes the calculations of medullary and cortical areas using this method. (B) Graph of total thymus area for each level. (C) Graph of K8⁺ cortical epithelial area for each level. (D) Graph of K5⁺ medullary epithelial area per level. (E) Three-dimensional representation of area across levels for total area, K8⁺ cortical area, K5⁺ medullary area. All graphs include standard error bars.