1. First of all, prepare a smear and let it air dry.
  2. Fix the smear with heat rolling the slide over blue flame for three times.
  3. Pour crystal violet over the smear and wait for 1 minute.
  4. Drain off excess stain and wash the stain with distilled water.
  5. Pour iodine over the stained smear and wait for 1 minute.
  6. Drain off excess iodine and wash the stain with distilled water.
  7. Pour 70% ethanol (15-30 seconds) or acetone (3-5 seconds).
  8. Pour Saffranin or dilute Carbol fuchsin and wait for 1 minute.
  9. Drain off excess stain and wash the stain with distilled water.
  10. Let the smear air dry and observe under oil immersion objective (100 x).

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