Jimmy Butler

Akash Mehta
4/17/19
Gram Staining lab report
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1. For each sample stained, state if it is Gram Positive or Gram negative, and describe the
shape/color. Attach a picture.
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Sample from cellphone:
—pink, therefore Gram negative
—scattering of dots; most are clustered together in large groups of dots; very few appear to be
standalone bacteria
—other parts (not captured in picture) look more purple, which may be a problem with our
staining or may be because there were more than one bacteria.

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Sample from keyboard:
—purple, therefore Gram positive
—much less concentrated than sample from cellphone

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—scattering of dots; often attached together as if strings of pearls, but also many standalone
bacteria
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2. Discuss the Gram Staining technique:
a. What is the role of crystal violet?
This is the die that, if it still appears at the end of the procedure, indicates that the
sample cells are Gram positive.
b. What is the role of iodine?
Iodine serves as a mordant, which is a chemical that fixes (i.e. binds) the dye,
keeping it in place.
c. What is the purpose of the alcohol wash?
It removes the dye and iodine from Gram negative (but not Gram positive) vells.
d. What is the role of safranin?
To make the Gram negative cells more visible.
e. Which bacterial structure is stained during this process?
The cell walls (and particularly the peptidoglycan layers).
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3. Describe the structure of the cell wall of Gram positive and Gram negative bacteria. Why
do Gram positive bacteria stain in purple while the Gram negative do not?
The Gram positive bacteria appear purple because in the staining/dying process,
certain chemicals (such as the iodine dye) will stick to Gram positive bacteria’s peptidoglycan
layer, but not to the Gram negative’s thinner peptidoglycan layer. Therefore, in subsequent steps,
the iodine will be washed away from the Gram negative bacteria, but will remain on the Gram
positive bacteria, keeping Gram positive purple and Gram negative pink.
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4. Discuss one reason why Gram Staining is helpful in the treatment of bacterial infections. Give
an example.
If doctors know whether a bacterial infection is Gram positive or negative, they
can prescribe antibodies that target only bacteria of that category.

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