University of the West Indies Mona

School of Nursing -Western Campus

Microbiology (NURS1111)

Laboratory Exercise # 2: Staining Total (20 marks)

Activity 1: The following organisms were gram stained read and report the gram reaction.
(6 marks)
 Species: Bacillus subtilis

Shape:

Arrangement:

Gram reaction

Species: Staphylococcus aureus

Shape:

Arrangement:

Gram reaction
 Species: Escherichia coli

Shape:

Arrangement:

Gram reaction

Activity 2

1. Explain the major differences between the gram-positive and gram-negative cell walls.
(2 marks)
2. Salmonella typhi is a gram-negative organism.
a. What color will it appear when simple stained with crystal violet? (1 mark)
b. What color would it be if it was gram stained correctly? (1 mark)
3. Explain how bacterial cells would look in the gram staining procedure if the following
mistakes were made: (3 marks)
a. Decolorizer left on too long
b. Decolorizer not left on long enough
c. Slide not heat-fixed before staining
4. What is the difference between a simple stain and a differential stain? (2 marks)
5. Explain why it is important to use only a small amount of bacteria when preparing a smear.

(2 marks)
6. What is the principle of the Gram stain? (2 marks)
7. State the names of two (2) simple stains. (1 mark)