New Gram Stain protocol

Hypothesis: Does methanol-fixing produce better gram-stain results than heat fixing?

1.)Wash slide with ethanol and lens paper; let dry
2.)Put bacteria on slide
3.)Let air dry
4.) NEW STEP * Cover bacteria/slide with methanol, let it sit for >2 minutes
5.)Pour excess methanol off, Let air dry…
Now continue the same way you would in previous method**
6.)Crystal Violet - 1 minute
7.)Rinse and blot
8.)Gram iodine - 1 minute
9.)Rinse and blot
10.)EtOH for decolorization; about 7-15 drops, use till it drips clear off slide.
11.)Rinse and blot
12.)Safernin - 1 minute
13.) Rinse and blot

*Cody Kruckenberg

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