Sample Lab Report – Prof.
Bissoon– Spring 2020
Exercise 3-6: Gram staining lab
For each lab report will need to include the following items as indicated in the lab report-
grading rubric:
Introduction and objective
Methods
Results
Conclusions
Gram Stain Lab Sample Lab Report – Exercise 3-6
Note: This is just a sample and should include as much detail as possible
for full credit!
1. Introduction: Explain the background of what you are doing and what you expect
to happen. This section should be detailed.
The purpose of this lab is to determine the morphology and Gram stain of various
bacteria under a microscope. The reason for staining bacteria is due to the fact that most
bacteria are transparent and cannot be seen through the microscope. The Gram stain is a
type of differential stain that allows a microbiologist to identify the differences between
organisms and/or differences within the same organism. The difference in staining is due
to the cell wall of Gram-positive and Gram-negative bacteria. Gram-positive cells have a
thick peptidoglycan cell wall, whereas, Gram-negative bacteria have an outer membrane
composed of various proteins and lipolysaccharides. Explain in details the stain’s that are
used in gram staining, as well as the bacteria being used.
1a. Objective: Must include the goal of the lab or the objective which is simply
stating what you did and why. The purpose of the Gram stain is to perform Gram stains
on various bacteria and to determine whether the bacteria are Gram-positive (purple) or
Gram-negative (pink).
2. Methods: Someone should be able to follow this and repeat exactly what you did.
Give as much detail as necessary to do this. If something was done for you by the lab
manager, or myself just say it in the methods section.
A drop of deionized water was placed on a microscope slide and the following bacteria
were put onto slides: Escherichia coli, Staphylococcus aureus, Micrococcus luteus and
Serratia marcescens. Then, the slide had to be air dried before heat fixing it, so that the
Gram staining may begin. The first step in Gram staining was to place the primary stain,
Crystal Violet, on the slide where the bacteria were heat fixed. The dye had to stay on the
slide for 30 seconds before rinsing with deionized water. Then the Mordant Iodine (which
is the Gram stain dye) was placed on the slide for another minute. After the minute and a
half was up, the slide was rinsed with deionized water. The next step was to use
isopropanol to wash off the dyes from the Gram-negative bacteria. To distinguish when
to stop using ethanol, the first drop of non-colored solution that slides off the slide was
the indicator. Again the slide was rinsed with deionized water and blotted dry. Lastly, the
saffarin dye was placed on the slide to counterstain the Gram-negative bacteria for one
minute, rinsed with deionized water, and blotted dry.
A light microscope was used to determine whether the bacteria were Gram-positive or
Gram-negative and cocci or bacilli.
3. Results: State your results here. Little explaining, just state the facts. However,
details are still important! Use pictures, charts, graphs etc….
Through the process of the experiment, E. coli was determined to be Gram-negative and
in the rod shaped or bacillus.
Then you would proceed with all your other results.
Pictures may be included here as well (be sure to label them properly).
Log sheets must be fully filled out and accurate if included for a lab.
4. Conclusions: This is where you want to give the most info. This is also the place to
discuss anything that may have gone wrong, troubleshooting, what you would do
differently to correct for anything that may not have worked correctly. You also
should try and connect this to the “real world” if possible. It is important to
understand what you did, why you did it and what the results mean.
Gram-positive and Gram-negative bacteria staining differ because of the composition of
their cell wall. Because the cell walls of Gram-negative cells have a higher content of
lipids and a thinner layer of peptidoglycan, the alcohol used in the decolorizing step made
the Gram-negative cells incapable of retaining the crystal violet stain. On the other hand,
Gram-positive cells have a thicker peptidoglycan that traps the crystal violet, making it
less vulnerable to decolorization.
The Gram stain technique was used on Escherichia coli, Staphylococcus aureus,
Micrococcus luteus and Serratia marcescens. E. coli was Gram-negative and was
bacillus shaped. Continue explaining the colors and shapes of the other bacteria tested.
Variation seen in this lab may be due to inappropriate staining such as the use of excess
primary and mordant iodine dyes being flushed out entirely by the 95% alcohol, or that
the mordant iodine dye was not left on long enough.
Then you should discuss why Gram-staining important? How it is used in the real world.
References: Properly list your references here if used in making your lab report.
Leboffe, M.J., Pierce, B.E. “Microbiology Laboratory Theory and Application”, Brief ed.
Moron Publishing Co. CO. 2008. p. 159-163.
