High Iron Diamine－Alcian Blue Stain
O-sulfate esters are stained brown-black with HID generated “in-situ” by the oxidation of a mixture of hydrochlorides of N, N-dimethyl meta- and para-phenylene diamines with ferric chloride. In subsequent steps, sialic acid and carboxyl groups are stained aquamarine blue with Alcian Blue at pH 2.5.
1. Alcian blue solution（pH2.5）
2. HID-solution A
3. HID-solution B
4. Ferric Chloride solution