The first test conducted was the Gram stain test. A single drop of saline was dropped onto a slide, after which a single colony was selected and immersified in saline then spread over the 1.5-centimeter slide. The slide was air dried, then heated and finally allowed to cool. Three to four drops of 1% crystal violet was added to the smear and left to stand for 30 seconds. Before the smear could dry, it was rinsed with distilled water and Gram’s iodine added. After 30 seconds, it was rinsed with 95% ethanol until the run-off was colorless. Safranin, was added and left to stand for 30 seconds. The slide was dried by gently blotting it on paper towel. It was examined under a compound microscope using 100X oil immersion objective to identify individual microorganisms.
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