Steps of gram staining procedure?

1. Prepare a heat-fixed smear of the bacterial sample on a clean glass slide.
2. Flood the smear with crystal violet stain and let it sit for 1 minute.
3. Rinse the slide with water to remove excess stain.
4. Flood the smear with iodine solution and let it sit for 1 minute.
5. Rinse the slide with water to remove excess iodine.
6. Decolorize the smear with alcohol or acetone for a few seconds.
7. Rinse the slide with water immediately after decolorization.
8. Counterstain the smear with safranin for 1 minute.
9. Rinse the slide with water to remove excess safranin.
10. Blot the slide dry with bibulous paper and observe under a microscope.

1. Preparation of the smear: A thin smear of the bacterial culture is made on a clean glass slide and allowed to air dry.
2. Heat fixation: The slide is gently heated over a flame to kill the bacteria and fix them to the slide.
3. Crystal violet staining: The slide is flooded with crystal violet solution and allowed to sit for 1 minute.
4. Washing: The slide is rinsed gently with water to remove excess crystal violet.
5. Lugol's iodine mordant: The slide is flooded with Lugol's iodine solution and allowed to sit for 1 minute.
6. Washing: The slide is rinsed gently with water to remove excess iodine.
7. Decolorization: The slide is flooded with 95% ethanol and allowed to sit for 10-30 seconds until the solution runs clear.
8. Washing: The slide is rinsed gently with water to remove excess ethanol.
9. Counterstaining: The slide is flooded with safranin solution and allowed to sit for 30 seconds.
10. Washing: The slide is rinsed gently with water to remove excess safranin.
11. Drying: The slide is allowed to air dry.