Staining Method of Hucker and Conn, Acetone, Grams Iodine, Carbol Fuchsin, Oil-immersion, Blot Dry, Ethanol Mixture

Staining Method of Hucker and Conn

	Staining Method of Hucker and Conn: Hucker and Conn in 1923 gave a modified method for Gram Staining (3), ‘is as follows. Prepare a smear and fix it by heat. Stain the smear with Hucker's crystal violet solution (823) for 1 minute. Add Gram's iodine (S24) to wash off crystal violet solution and allow the smear to react with fresh iodine for 1 minute. Rinse the slide in running water.
Add decolourizing solution (acetone and ethanol mixture, 1:1) drop wise at the upper end of slide held in inclined position, till the washing does not contain violet colour. For normal smear 10-15 seconds are enough. Rinse the smear with water. Counterstain with safranin (S25) or 10 times diluted carbol fuchsin (S19) for 30-60 seconds. Blot dry and examine under the oil-immersion objective.