The mechanism of bilateral staining of the cortex of wool fibers by basic dyes has been investigated in detail. Merino wool fibers were treated with formic acid and pronase, and their behavior in staining with basic dyes such as methylene blue and janus green was examined using light and electron microscopy. Formic acid is known to remove intercellular cement, one of the nonkeratinous proteins, from the cell membrane complex, while pronase removes all nonkeratinous proteins. Bilateral staining was still distinctly observed after cuticle removal, but not for wool fibers treated with pronase, for fiber fragments obtained by treatment with formic acid with stirring, or for cortical cell particles recovered after fibrillation of the cortex in formic acid. The microfibril-matrix structure of the cortex remained unchanged, however, even after treatment with formic acid and with pronase. These results imply that bilateral staining with basic dyes occurs because of the difference in the network structure of nonkeratinous proteins, especially the intercellular cement of the cell membrane complex between the orthocortex and paracortex.The cortex of fine wool fibers consists of two major segments, the orthocortex and the paracortex, and the bilateral arrangement of these two cortices is closely associated with the crimped structure of wool fibers [2,3,12]. Since its discovery by Horio and Kondo [9] and Mercer [ 18], many researchers have studied the differences in the responses of each component of the bilateral cortex to various dyes and metal ions [2]. These differences have been interpreted in terms of differences in the fine structure or the reactivity between orthocortex and paracortex [2,4,9,10,14,18,19,21 ]. There are several possible differences between the two cortices, however, and their behavior in staining is different with ,dyes and metal ions [2]. The detailed mechanism of bilateral staining of the cortex is still open for discussion to some extent. The pioneer works by Horio et al. [9,10] were done with ionic dyes. The staining mechanism with dyes is relatively simple compared to that with metal ions, which is specific to individual ions. A possible interpretation is that the content of charged groups is different for the orthocortex and paracortex. Wool fibers are composed of a number of ceU components, however, and the accessibility of dye molecules to the charged groups is different for different cell components. There is insufficient experimental evidence to explain why large dye molecules can produce a remarkable contrast in staining between the two cortical components.In a previous paper [ 11 ], we reported on the amino acid composition of the isolated orthocortical and paracortical cells. The results showed that there is little difference in the relative content of the charged groups except for cystine. During the course of this study, we also found that bilateral staining does not occur alter fibrillation of the cortex. These results suggest that the nonkeratinous materials [2,24], which may be remove...