Studies on Papanicolaou staining. III. Quantitative investigations of orangeophilia and cyanophilia.

This paper provides data derived from the visible light absorbance spectra of Papanicolaou stained epithelial cells from the uterine cervix. Twenty-four types of spectra have been considered, namely, those derived from orangeophilic and cyanophilic nuclei and cytoplasms of superficial, intermediate, parabasal and dysplastic cells, and cells of carcinoma in situ and invasive carcinoma. Wavelengths of maximum absorbance and peak absorbances are tabulated. The proportions of bound orange G, eosin Y, aluminum-hematein and light green SF yellowish have been calculated. For the majority of cell types, dyebinding differences between orangeophilic and corresponding cyanophilic substrates were statistically significant. CIE coordinates were calculated from absorbance spectra; again differences between organeophilic and cyanophilic cells were statistically significant in most cases. Although the designation of cells as orangeophilic or cyanophilic is made on the basis of cytoplasmic coloration, the nucleus is also usually orangeophilic or cyanophilic. These nuclear differences are real and not due to the effects of over- and underlying cytoplasm.