MICROBIOLOGY LABORATORY ACTIVITY NO.

GRAM STAINING: A VIRTUAL LABORATORY EXERCISE

LABORATORY REPORT

Name: Blethy April C. Palao Rating: _____________

Course/Block: BSED 2-1 Date: 10 / 08 / 2021

IV. DATA AND ILLUSTRATIONS

1. List down the equipment, substance/chemicals used based on the virtual laboratory.
LABORATORY EQUIPMENT SUBSTANCES/CHEMICALS

1. Glass Slide 1. Crystal Violet

2. Bunsen Burner 2. Iodine

3. Microscope 3. Alcohol

4. Wash Bottle 4. Safranin

5. Water (H2O)

2. Attach a screen recording or video of you performing the laboratory activity together with
the laboratory report. Length of screen recording/video must not exceed 3minutes.

V. RESULTS AND DISCUSSION

Directions: Answer each question in not more than five sentences.

1. Describe the function of each of the following in the Gram Staining process:

a. Crystal Violet – It serves as the primary stain or the basic dye for the cells. At this
point, both the gram negative and gram positive cells will appear purple when viewed
in a microscope. This is because the basic dye is positively charged, and it adheres
to the cell membrane of the gram negative and gram positive cells.

b. Iodine – It is used to fix the crystal violet on the bacterial cell wall. Aside from this,
Iodine is also used to combine with the crystal violet in order to form a di-iodine
complex. This is used to decrease the solubility within the cell.

c. Alcohol – It is used to sort the gram positive cells from the gram negative cells by
decolorizing them. The gram-positive bacteria retain the crystal violet. However, the
gram-negative bacteria do not.

d. Safranin – The safranin is used to stain the decolorized gram negative cells in order
to separate it from the gram positive cells.

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 2. Which step in the Gram stain is most likely to cause poor results if done incorrectly?
The most critical step in the gram staining process is the differential step or the
decolorization step. If the decolorization process is done excessively, the crystal violet
stain will be removed on both gram positive and gram negative cells. In that case, the
gram positive might also appear as gram negative. This will cause confusion and poor
result if this step is done incorrectly.

3. Why is Gram staining an important diagnostic tool used in treating patients with
suspected infections?
Through the process of gram staining, the bacteria in a particular sample can be
characterized depending on the stain. And with that, gram staining will help to determine
if the patient has a bacterial infection. Gram staining is also important in order to
determine what type of bacteria are causing the infection. Through this, it will be easier
to assess the type of medication that the patient should take.

VI. CONCLUSION

Directions: Generate a conclusion based on your experience in performing the virtual

laboratory activity and the importance of Gram staining in not more than five sentences.

The gram staining is the most widely employed staining method in microbiology
because it divides bacteria into two classes, gram positive and gram negative. Gram
staining has different steps involving heat fixing, primary staining using crystal violet,
fixing the dye using iodine, decolorizing using alcohol, and counterstaining using safranin.
Each step should be done properly and with caution since it will cause a major effect on
the result. The process of gram staining is very important since it is used in diagnosing
and treating patients with bacterial infections.
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