Gram Staining Lab Supplies: 1. Bacterial culture 2. Gram stain kit 3. Slide 4. Metal loop 5. Bunsen burner 6.

Microscope Procedure: 1. Begin by observing the culture plates. What are the differences between the before and after plates? How many colonies of bacteria are on each plate? Do they look any different? 2. Choose a plate that has an isolated colony of bacteria on it. Turn on your Bunsen burner as demonstrated. Flame you loop and then touch an open part of your agar to cool it. Then swipe the loop through your chosen colony of bacteria. Apply the loop to a drop of sterile water on your slide. Pass the slide through the flame of the Bunsen burner to heat fix the slide. Flame your loop again and extinguish the flame. 3. Look at your slide under the microscope as demonstrated earlier. Record your observations. 4. Gram stain your slide as follows: a. Apply the crystal violet stain to your slide. After 30 sec rinse with water. b. Apply the iodine solution (30 sec) and rinse again. c. Rinse quickly with alcohol (until water runs clear). d. Apply fuschin dye (30 sec) and rinse. e. Observe under a microscope and record your findings. 5. At the end of the lab be sure to properly dispose of all plates and slides. Put the microscopes back in the proper cabinet. Make sure you have turned the gas off!!!

1. Observations of plates:

2. Observations of slide before staining:

3. Observation of slide after staining:

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