presentation JTC 300

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By Nichole Guidotti:

By Nichole Guidotti

Performing a Gram stain:

Performing a Gram stain

Step one: preparing the slide:

Step one: preparing the slide Obtain a culture of bacteria from an agar plate. Sterilize your inoculating loop and wait 30 seconds. With the inoculating loop, obtain a small amount of bacteria from the agar plate and place on a microscope slide Add a drop of water to the slide and wait for it to dry Heat fix the slide by passing through the bunsen burner

Primary stain: Chrystal violet:

Primary stain: Chrystal violet 1. Add enough Chrystal Violet to cover the slide 2. Wait one minute 3. Wash off stain with water

Mordant: Grams Iodine:

Mordant: Grams Iodine 1. Add enough Grams Iodine to cover the slide 2. Wait one minute 3. Wash off slide with water

Decolorizer: Acid Alcohol:

Decolorizer : Acid Alcohol Place decolorizer on slide one drop at a time, not exceeding 30 seconds Wash off with water

Secondary stain/ Safranin:

Secondary stain/ Safranin Add enough Safranin to cover the slide Wait two minutes Wash off with water View slide under microscope


Purpose: -Gram Stains are the first step to differentiating between Gram positive and Gram negative bacteria -Gram negative bacteria stain pink while Gram positive bacteria stain purple.

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