Name: _______________________________________________ Lab Section: ____________________

Gram Stain Report

This written report is worth 15 points.
Assignment What do you need
Each student will have the opportunity to submit one of your Gram stains to turn in?
with a brief written report describing the staining procedure and the stain
A microscope slide of
itself. A good report will necessitate a mini-mum of 2 double-spaced pages
and is limited to a 3-page maximum.
your best Gram stain or
I. Overall Format a photograph with
Please include the following in your report: detailed observation
1. Title Page with the following information: A 2-3 page report
A. Title: A title should be brief, creative, but descriptive of (double-spaced) about
the work that was done / findings. the slide submitted
For tips on writing a
B. Your Name good scientific report,
C. Course and Lab Section see Page xxxi.
D. Date
(1 point) __________
2. Introduction:
An introduction should be written as a funnel. That is, it should present the broad significance first.
Indicate why this topic is important to absolutely every reader. Include pertinent background information.
In this case, the history of the Gram stain and its significance in unknown identification are relevant. A
discussion of the mechanism of Gram staining and how it differentiates bacteria on the basis of their cell
wall structure should also be included (2 points for broad significance / background). The Introduction
should then funnel to the narrower purpose and objectives of the experiment. Finally, at the bottom of the
funnel, present the hypothesis. In this case, based upon background knowledge, a hypothesis as to whether
the unknown will be Gram-positive or Gram-negative can be made. Remember that a hypothesis need not
be correct, but it must be tested by the procedure (2 points for narrowing the funnel and presenting a
purpose, objective and hypothesis). (1 point for focusing and unifying - a good Introduction keeps the reader
focused on the overall goal and shows connection between all sentences.)
•Hypothesis Clarification (The hypothesis for this experiment will the trivial. That is, prior
experiments have already determined the Gram reaction of your unknown. However, this will be the
first time that you have determined the Gram reaction for an unknown and, as such, you will likely
have an educated guess as to whether your unknown is Gram-positive or Gram-negative. Write this
guess as a hypothesis. Please use one of the two following formats.
a. Manipulated hypotheses: This type of hypothesis is used when one variable (the independent
variable) is manipulated (usually by the experimenter) and the effects on a second variable (the
dependent variable) are observed. This type of hypothesis is generally written as an “if, then”
statement (e.g. If the smear of the unknown bacterium is decolorized as according to the above procedure (neither
over- nor under-decolorized), then all the cells will stain pink (Gram-negative.) In this example, decolorization is
the independent variable (the variable controlled by the experimenter) and the color of the cells is
the dependent variable (the variable affected by the changes in the independent variable). While the
above example is trivial, it is testable and making such a statement enables practice with scientific
reasoning. Another example of how an if, then statement might be made is as follows: If the unknown
bacterium is found to be a coccus, then it will stain Gram-positive. In this example, a quality of the unknown is
the independent variable. The dependent variable relies on this quality.
b. Observational, or predictive, hypotheses: This type of hypothesis makes a prediction about that
which will be observed during the procedure (e.g. The unknown bacterium will stain Gram-
positive.). Note that, like the first example, this hypothesis can be tested by the procedure.
(4 points) __________
■ General and Medical Microbiology Lab 155
 3. Materials and Methods:
This section should include a description of the procedure/steps in the Gram stain, including the
preparation of a bacterial smear and heat fixation. This section must be written in complete sentences and in
paragraph form (1 point). Illustrations may assist in clarifying points. It is often easy to confuse the Materials
and Methods section as being synonymous with the procedure written in the lab manual. However, the
Materials and Methods section relates exactly what was done rather than relating a plan (1 point). This section
should enable a reader to repeat your steps verbatim (1 point). For example, an excerpt from the Materials and
Methods might read: The slide was decolorized with ethanol for just long enough to enable the runoff to change from purple to
colorless. This took approximately 25 seconds. Immediately after the change in runoff color, water was run over the slide using a
squirt bottle. This was done for approximately 30 seconds; the slide was then flicked repeatedly to remove excess water...
(3 points) __________
4. Results:
In this section, in your own words, thoroughly describe what you observed about the unknown Gram stain.
Include at least one illustration or detailed photograph of the Gram stain (1 point). Descriptive observations are
the cornerstone of science and your illustration and description must reflect the actual appearance of your slide
(1 point). An excerpt of the observation may read something like, The unknown Gram-stain was too thick as evidenced
by areas where bacteria were piled so high that there were multiple colors. However, in all areas where the smear was of appropriate
thickness, the cells stained pink. It was difficult to discern any regular arrangement as there were some areas where cells were back-
to-back but in most areas they simply seemed scattered...
(2 points) __________
5. Discussion:
A discussion is very different from a Results section. Whereas a Results section is the section in which you
relate exactly what you saw or measured, the Discussion is where you make sense of it. That is, the Discussion
is a place for interpretation, a place to make meaning of your results.
In the case of this experiment, discuss what you now know about your unknown and what it tells you about
its cell wall. Indicate how this will guide your further work to identify the unknown. Were there features of the
Gram stain that are not as expected? What possible sources of error may be responsible (e.g. poor quality of the
smear preparation)? Discuss whether the hypothesis made in the introduction was tested and, if so, was it
supported or rejected/modified?
(3 points) __________
II. Overall Quality
Students have the opportunity to earn points for reports that are well written, include proper citation and are
organized and edited. Reports that lack citation, are not edited or do not follow the structure described above
may be returned to students in order to enable them to earn these points. If the report quality is so low that it
causes TAs to spend inordinate amount of time trying to interpret it, and if the student chooses not to rewrite
the report for higher quality, than the TAs have the prevue to deduct 1-2 additional points from the overall
grade.
(2 points) __________
DUE DATE: ________________________ (Your TAs will give you this date.
TA’s suggestions for improving Gram stain: _____________________________________________
_______________________________________________________________________________
_______________________________________________________________________________
Overall TA Comments: ____________________________________________________________
______________________________________________________________________________
_______________________________________________________________________________
Please tear this grading key out of your lab book and attach it to your lab report at the time of
submission. Please also write your name and lab section on the frosted portion of your Gram
stain slide (if you are submitting this in place of a photograph and detailed observation) and
hand it in with your written report.

Points earned out of 15 possible: ____________

156 ■ General and Medical Microbiology Lab