Kyle Hammermeister MT 213 A37412503 Gram/Stain Tutor 1) Step 1- Flood the slide with the crystal violet reagent for one minute then rinse with water Step 2- Flood the slide with the gram’s iodine reagent for one minute and rinse with water. Step 3- Decolorize the slide by gently rinsing with acetone-alcohol solution for ten seconds then rinse with water. Step 4- Flood the side with the reagent safranin for one minute and rinse with water and let air dry. If the gram stain is positive for bacteria and yeast the stain will be dark purple. If the gram stain is negative for bacteria and host cells it will be stained pink. 2) The characteristics that determine if a gram stain is positive or negative include the chemical and physical properties of the bacterias cell walls. Since the color depends on the permiablility of the cell wall a gram negative cell wall will have more
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