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Gram stain procedure


This stain for differentiate between G +ve bacteria (which accept staining) and G -ve bacteria (which not accept staining).
Steps:
-Prepare your clear slide and put one drop of normal saline.
-Take specimen from the colony and mix it well with normal saline.
-Leave for dryness on air then fix it on burner.
-Then pour Crystal Violet stain on the slide and leave it for 1 minute.
-Wash gently with water.
-Then pour Iodine stain on the slide and leave it for 1 minute.
-Wash gently with water.
-Wash with Alcohol to decolorize the slide.
-Then pour Safranin stain on the slide and leave it for 1 minute.

Results under microscope:
-Violet cells refer to Gram Positive bacteria (G+ve) and red cells which stained by Safranin refer to Gram Negative bacteria (G-ve).

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