Gram-Positive or Gram-Negative

Hiba Khaled Al Kurdi

ID: 202100288

Teaching Science in Highschool

Instructor: Dr. Salam Noureddine
Fall Semester 2022-2023
Gram Staining:
 A bacteriologist named Hans Christian Gram attempted to discover a universal
stain that would work with all bacteria. Thus, he discovered that all bacteria
could be either gram +ve (retain stain) or gram –ve (do not retain stain).
 Gram +ve or –ve differ in the characteristics of their cell wall mainly, but this
does not determine “good” or “bad”. However, distinguishing if a bacteria is
gram +ve or –ve could direct doctors in diagnosis since different bacteria cause
different diseases.
 For example: bacteria causing scarlet fever is gram +ve, and that causing
typhoid or cholera is gram –ve.

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To Prove This:
1. Experimenting:
 Collect the sample by scraping tissue from the
surface of a part of your body (for eg: throat, skin
wound..)
 On a clean slide spread or smear the sample to be
put under the microscope.
 Apply the primary stain: crystal violet
Click icon to add picture
 Add a mordant: iodine
 Decolorize with ethanol and acetone (95%)
 Apply a counterstain: safranin

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2. Observing:
 Upon adding the primary stain, the all the bacteria present were stained violet.
 Then a mordant (iodine) was added to fix the color absorbed by the sample.
 However, when ethanol (alcohol) was added at a certain percentage to wash the
primary color despite the presence of a mordant, the stain on some was not retained
and it vanished.
 Once the counterstain (sufranin having a pink color) was added, the bacteria
(organisms under the microscope) that did not retain the primary stain turned pink
while the ones that did remained violet (initial color).

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 VS

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3. Predicting – Hypothesis:
 When staining bacterial cells, the stain should cross the cell wall to enter and color the
cells. Thus, the fixation of the mordant and the decolorization of the stain are related to
the composition of the cell wall.
 Based on our known info that cell walls are mainly composed of peptidoglycans, so the
results show that there is a difference in the molecular weights of peptidoglycans
between gram +ve and gram –ve. Also, since one type didn’t retain the stain so it might
be considered as a negative test and its negative test could be a result of less
peptidoglycan amounts than the other one.
 Hypothesis: Gram negative gave the pink results and gram positive gave the violet
results.

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4. Analyzing & Inferring:
 Gram –ve bacteria contain an additional membrane (outer membrane)
than the gram +ve which is the LPS. So, during gram staining, the
outer membrane deteriorates from the alcohol added to the sample and
since it has a thin layer of peptidoglycan, it will not be able to retain
the crystal violet stain. Thus, it takes the color of Sufranin added lastly.
 While gram +ve since it has a thicker peptidoglycan cell wall that is
much more stable, so it will retain the initial stain color despite the
added alcohol.

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5. Communicating More Infos:
 Although gram –ve bacteria have a thinner cell wall
composition of peptidoglycans, yet it is much more resistant
to antibiotics than gram +ve bacteria.
 This is due to most antibiotics target the cell wall in order to
kill bacteria but in gram –ve the peptidoglycan is so much
thin thus it can not be recognized by the antibiotics, making
it very resilient.
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 GRAM POSITIVE GRAM
NEGATIVE

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 Click icon to add picture

Thank you!
Hope you enjoyed
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