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