Gram Staining

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Gram Staining

• Lab 3

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Hans Christian Gram

• Developed
• Gram Staining
• in 1844

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The reagents

• Primary stain Crystal violet
• Mordant Iodine
• Selective TreatmentEthanol
• Counterstain Safranin

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To Gram stain a smear

• 1. Flood the slide with crystal violet and let it
  stand for 30 seconds.

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To Gram stain a smear

• 2. Remove the crystal violet by rinsing gently
  with iodine.

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To Gram stain a smear

• 3. Flood the smear with Grams iodine and allow
  the iodine to react for about 2 minutes

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To Gram stain a smear

• 4. Decolorization step Rinse with ethanol until
  color stops following then rinse with water.

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To Gram stain a smear

• 5. Apply the counterstain, safranin for 3-5
  minutes and then rinse stain from slide.

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