Manipulating the Papanicolaou staining method. Role of acidity in the EA counterstain.

The role of phosphotungstic acid as the main pH-controlling agent in Papanicolaou's EA stain was analyzed, including its effect on the competition between eosin Y and light green, the principal components of EA solutions, and this acid was compared with HCl as an acidifying agent. The amount of phosphotungstic acid in the solution determined the color of the cytoplasm. Our study led to a better understanding and a simplification of the staining procedure. With the acquired knowledge, the Papanicolaou counterstain can be standardized to suit the taste of the individual cytologist and can add to the predictability of the results.