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The Gram Stain Procedure 7 лет назад


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The Gram Stain Procedure

The Gram Stain Procedure. This protocol is taken from "Benson's Microbiological Applications: Laboratory Manual in General Microbiology" Eleventh Edition. Some protocols vary depending on the source. Use the one best suited for your laboratory. The gram stain follows this procedure: 1. Prep your sample on your slide 2. Heat fix the slide 3. Flood with Crystal Violet for 20 seconds 4. Wash for 2 seconds 5. Flood with Gram's Iodine for 1 minute 6. Decolorize with Grams Alcohol for 10-20 seconds 7. Wash for 2 seconds 8. Flood with Safranin for 1 minute 9.Wash for 2 seconds 10. Blot the slide dry Gram-Positive bacteria have a thick peptidoglycan layer on their cell membrane which retains the crystal violet-iodine complex and stains the bacteria purple. Gram-Negative Bacteria have a thin peptidoglycan layer on their cell membrane which does not retains the crystal violet-iodine complex and is instead stained by the Safranin. This will leave the cells stained pink.

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