Alcian Blue reacts to compounds containing anionic groups such as acid mucosubstances and acid mucins. At pH 2.5, both carboxylated and sulfated acid mucosubstances are stained blue. Periodic acid is used to oxidize the tissue to form aldehyde groups that are demonstrated by Schiff Reagent. Glycogen and neutral mucosubstances as well as basement membranes will be stained magenta by Schiff Reagent, followed by a tap water rinse to develop and intensify its magenta hues. An optional counterstain may be used to provide nuclear detail. Hematoxylin 7211 is recommended; however, Kernechtrot Nuclear Fast Red may be used if a red nuclear stain is desired

Fungal cell walls are rich in polysaccharides known as glucans. Periodic Acid Solution oxidizes these polysaccharides to form aldehydes, which in turn react with the silver ions present in the Working Methenamine Silver Solution and reduce them to form visible metallic silver. Gold Chloride Solution tones the sections and intensifies the reduced silver by forming a silver-gold complex. Sodium Thiosulfate Solution removes any unreduced silver from the tissue sections, followed by Fast Green Stain Solution which produces a light green background to further enhance contrast.