Virtual Lab Report by Honey

Noreen Belnas

The Gram Stain: Identify and differentiate

bacteria
This lab report is for you to reflect on what you completed and learned in this
simulation, and to practice your written scientific communication skills.

Sections

1. Describe the overall objective and make a hypothesis

2. Introduce relevant background knowledge on this topic
3. Summarize the steps taken in the simulation
4. Explain any obtained results
5. Discuss the conclusions and implications

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 1. Describe the overall objective and make a hypothesis.
What is the overall purpose of the experiments or activities? Make a hypothesis if applicable.

Hint: The purpose is often stated in the welcome message of the simulation.

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As we perform the simulation, the following objectives are:

 Describe the structure of the Gram-positive and Gram-negative bacteria.

 Appreciate theoretical and technical aspects of the Gram staining procedure.

 Know the most commonly made mistakes in Gram staining.

 Critically interpret the results of a Gram staining experiment using a light

microscope.

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 2. Introduce relevant background knowledge on this topic.
What have you learned in class or researched on your own that would help prepare for this
simulation?

Hint: You can review the “THEORY” section in the simulation or at https://theory.labster.com/
if you need help.

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The relevant knowledge that we gained as we perform the simulation are:

Exploring the bacterial cell wall

We compare and contrast the cell wall of Gram-positive and Gram-negative bacteria by

building your very own bacterial 3D models on the hologram table. We also enter to the

exploration pod to observe in an immersive animation how the four reagents of the Gram

stain interact with structural components of the cell wall to color the bacteria.

Performing the Gram stain

When the patient’s fluid sample arrives at the laboratory, we equip ourselves virtually with

protective gear to prepare a bacterial smear and heat fixes it to a glass slide. We are now

ready to perform the Gram stain in a safe virtual environment. If we accidentally made a

mistake, we can hit the big red button on the workbench to repeat the staining procedure

until it becomes second nature.

Interpreting our findings using a microscope

In the end, we used a light microscope to interpret the results of your Gram stain. We view

the microscopic image on the computer screen, and apply immersion oil to increase

magnification 1000x!

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 3. Summarize the steps taken in the simulation.
Explain each step you completed including the equipment and techniques you used.

Hint: You can use the “MISSION” tab in the LabPad as inspiration.

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First, we get ready to work on the lab, a lab coat is essential in going to the lab. As soon as

we enter the lab, let’s also not forget to wear our gloves. Next, we visit the hospital and

identify the protection of Central Nervous System (CNS) and then we equip ourselves to

work in the lab safety. As we go along in our simulation, we were presented a bacterial cell

structure and its 3D model. In the simulation the gram stain animation was also presented.

We apply smear and heat-fix sample, and perform the gram stain until the first wash,

second wash, third wash, and completion. After that, we also go to the Microscopy bench

where we play a mini-game.

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 4. Describe any obtained results.
Explain any obtained results. Were these results expected or unexpected?

Hint: You can use the “MEDIA” tab in the Lab Pad to find relevant images from the
simulation. You can also take screenshots while you are playing the simulation.

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Using a sterile inoculating loop, add 1 drop of sterile water to the slide. Prepare a mixed

smear.

2. Air dry and Heat fix.

3. Cover the smear with Crystal Violet (primary stain) for 1 min.

4. Gently wash off the slide with water.

5. Add Gram’s Iodine (mordant) for 1 min.

6. Wash with water.

7. Decolorize with 95% ethanol. Stop decolorizing with alcohol as soon as the purple color

has stopped leaching off the slide (time will vary depending on thickness of smear).

Immediately wash with water. Be sure to dispose of all ethanol waste in the appropriately

labeled waste container.

8. Cover the smear with Safranin for 30 seconds.

9. Wash both the top & the bottom of the slide with water.

10. Using the 10X objective lens, focus first on the line and then on the smear. Follow the

focusing procedure in Lab #1. Using the focusing procedure in Lab #2 to view the smear

using the 100X (oil immersion lens).

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 5. Discuss the conclusions and implications.
How do your results relate back to the original purpose and your hypothesis? Were there any
systematic sources of error that could have affected the results? What did you learn? What
is the importance of these findings and how can you apply them to other real world
situations?

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Interpret our findings using a microscope

In the end, we used a light microscope to interpret the results of your Gram stain. We view

the microscopic image on the computer screen, and apply immersion oil to increase

magnification 1000x! If your staining procedure was successful you will see Gram positive

cocci

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