GRAMS IODINE FOR MICROSCOPICAL STAINING

1. Add 20mg safranin powder to a 100ml beaker.
2. Pour 20ml distilled water in the beaker and make 0.1% safranin staining solution by constant stirring.
3. Transfer 20mg of fast green dye in another 100ml beaker. ...
4. Filter both the staining solutions to avoid particles.

How to use Gram's Iodine Solution

1. Prepare a thin smear on clear, dry glass slide.

2. Allow it to air dry and fix by gentle heat.

3. Flood with Gram's Crystal Violet (S012) for 1 minute. (If over staining results in improper decolourization of known gram-negative organisms, use less crystal violet).

4. Wash with tap water.

5. Flood the smear with Gram’s Iodine (S013). Allow it to remain for 1 minute.

6. Decolourize with Gram's Decolourizer (S032) until the blue dye no longer flows from the smear. (Acetone may be used as a decolourizing agent with caution, since this solvent very rapidly decolourized the smear).

7. Wash with tap water.

8. Counter stain with 0.5% w/v Safranin (S027) for 20 seconds and rinses off with water.

9. Wash with tap water.

10. Allow the slide to air dry or blot dry between sheets of clean bibulous paper and examine under oil immersion objective