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Laboratory Result

This document describes the results of a gram staining laboratory experiment. [I] It instructs the student to draw gram stained microorganisms and label their morphology. [II] The results table identifies various microbes' morphology, gram staining reaction, and whether they are gram positive or negative. [III] It asks the student to identify the primary stain used (crystal violet), describe the function of mordant, primary stain, decolorizer and counterstain, and explain that the decolorization step is most crucial as incorrect performance could yield inaccurate gram staining results.

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Sean Sarigumba
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226 views5 pages

Laboratory Result

This document describes the results of a gram staining laboratory experiment. [I] It instructs the student to draw gram stained microorganisms and label their morphology. [II] The results table identifies various microbes' morphology, gram staining reaction, and whether they are gram positive or negative. [III] It asks the student to identify the primary stain used (crystal violet), describe the function of mordant, primary stain, decolorizer and counterstain, and explain that the decolorization step is most crucial as incorrect performance could yield inaccurate gram staining results.

Uploaded by

Sean Sarigumba
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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Laboratory Result

I. Draw the microorganisms seen after performing gram staining. Then label the
bacterial morphology (shapes and arrangements (4 points).
II. Result from Gram Staining. Fill in the table below:

Microbe Morphology Gram Staining


Reaction
Staphylococcus aureus Staphylococcus-round Gram positive
shaped and in grape bacteria
like clusters
Pseudomonas aeruginosa Bacilli-slender and rod Gram- negative
shaped, asporogenous, bacteria
and monoflagellated
bacterium
Streptococcus pneumoniae Streptococci- lancet- Gram positive
shaped cocci (elongated bacteria
cocci with a slightly
pointed outer
curvature). Usually, they
are seen as pairs of
cocci (diplococci), but
they may also occur
singly and in short
chains.
Corynebacterium diptheriae Bacilli-slender, usually Gram positive
with one end being bacteria
wider, thus giving the
often-described club-
shaped appearance.
Haemophilus influenzae Bacilli or coccobacilli- Grame negative
nonmotile rod shaped bacteria
bacterium that are
small, pleomorphic with
random arrangements
Streptococcus pyogenes Streptococci- round to Gram positive
ovoid coccus, chainlike bacteria
shape from
interconnected spheres
Neisseria gonorrhoeae Neiserriae- kidney bean Gram negative
shaped cocci, 0.6 to 1.0 bacteria
µm in diameter, usually
seen in pairs with
adjacent flattened sides
Bacillus anthracis Bacilli- rectangular, Gram positive
aerobic, rod-shaped bacteria
bacterium with square
ends, measuring about
1µ x 3-5µ with a
common chain
formation

III. Answer the questions below : 


1. Which stain is the primary stain for the Gram stain? ( 1point)
The Gram staining method is one of the most important staining techniques in
microbiology. It is almost always the first test performed for the identification of
bacteria. In this regard, the primary stain used is the crystal violet stain. Crystal violet is
sometimes substituted with methylene blue, which is equally effective. The
microorganisms that retain the crystal violet-iodine complex appear purple brown under
microscopic examination. 
2. Describe the function of each of the following in the Gram stain ( 1point each)
Mordant: a mordant is a compound used to hold down molecules of a stain
onto a microorganism. It is also used to form crystal violet iodine complex in
cell, mostly in gram positive bacterial cell because of thick peptidoglycan
layer and it increases the affinity for crystal violet in cell.

Primary Stain: The primary stain (crystal violet) binds to peptidoglycan,


coloring cells purple. After a sample of bacteria on the glass slide is heated,
primary stain is applied to the sample. Both gram-positive and gram-negative
cells have peptidoglycan in their cell walls, so initially, all bacteria stain violet.
Decolorizer: This dehydrates the peptidoglycan layer, shrinking and
tightening it. The large crystal violet-iodine complex is not able to penetrate
this tightened peptidoglycan layer and is thus trapped in the cell in Gram
positive bacteria. Conversely, the outer membrane of Gram-negative bacteria
is degraded and the thinner peptidoglycan layer of Gram-negative cells is
unable to retain the crystal violet-iodine complex and the color is lost.
Counterstain: It is the final step in gram staining and is applied to the smear
to give decolorized gram-negative bacteria a pink color.
3. Briefly describe the mechanism of Gram staining. ( 2 points )

Gram staining is a common technique used to differentiate two large groups of bacteria
based on their different cell wall constituents. It involves three processes: staining with a
water-soluble dye called crystal violet, decolorization, and counterstaining, usually with
safranin. The Gram stain procedure distinguishes between Gram positive and Gram-
negative groups by coloring these cells red or violet. Gram positive bacteria stain violet
due to the presence of a thick layer of peptidoglycan in their cell walls, which retains the
crystal violet these cells are stained with. Alternatively, Gram negative bacteria stain
red, which is attributed to a thinner peptidoglycan wall, which does not retain the crystal
violet during the decoloring process.

3. Which step in the Gram stain is most likely to cause poor results if done
incorrectly? Why? ( 3 points )  
The decolorization step is the most crucial step in Gram staining is most likely to
cause poor results if done incorrectly. It should be performed very carefully
because the decolorization agent is applied for too long, the Crystal violet
stain will be removed from the Gram positive cells making them appear Gram
Negative whereas Under-decolorization will result in Gram Negative cells appear
as Gram positive.

Post Virtual Laboratory Reflection 

After gram staining, answer each question thoroughly, including evidence to


support your claims. Your response must be well thought out and written in a coherent
manner. Respond concisely to less than 300 words.

1. The most interesting parts of this virtual lab experience were:

What I found most interesting during this virtual lab experience was the way I grew to
understand the vital role that gram staining had within the clinical setting. When
examining a smear of, say, pus from an abscess, this stain often allows to include for
consideration roughly half the clinically relevant bacterial species while excluding the
others. Or consider a patient with meningitis. Here, speed is of the essence because
treatment must be initiated right away. A Gram stain of the spinal fluid may reveal within
minutes the presence of Gram-positive cocci, , Gram-negative cocci, or Gram-negative
slender rods .This can make a critical difference in the choice of antibiotics that have to
be administered in great haste. It  is cheap, effective, quick, and relatively easy to
interpret and goes a long way in helping nurses to make a treatment plan and the
appropriate nursing interventions

2. The hardest part of this virtual lab experience was…because…

For me, the hardest part of this virtual lab experience was having to visualize the entire
procedure instead of doing it within the premises of a laboratory. This is because I
consider myself to be more of a visual learner and since this is a topic which is often
based on practice, being unable to perform the actual procedure places limitations on
what we can actually learn compared if we were to perform it firsthand.

3. The one thing I learned and that I think will stick with me is

The one thing I learned and that I think will stick with me is that I need to be especially
careful during the decolorization phase of gram staining because the entire Gram
staining procedure hinges on the successful decolorization with ethyl alcohol step is not
only difficult, but it will throw off your results entirely if it is done incorrectly. If you do it
for too long, you'll get false Gram negative organisms and if you under-decolorize, you'll
get false Gram positive organisms.

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