EXERCISE NO.

6
PREPARATION OF
SMEARS AND SIMPLE
STAINING GROUP 5

Jinang
Sanday
Gellor
Malambut
Tuanzi
Bonao
GROUP 5 APRIL 26, 2021

Bacteria are so minute and almost transparent. The slight

difference between the refractive index of live organisms and
water makes observations of live cells difficult under the bright
microscope, especially if the cells are motile. To observe forms
and structures of bacteria better, some pre-treatments are
necessary to immobilize the cells and to differentiate their
structures. These include fixing and staining bacteria in a smear
preparation.
GROUP 5 APRIL 26, 2021

Objectives:

1 To prepare a good smear. 3 To differentiate the different forms of

bacteria.

2 To perform a simple direct stain. 4 To list the advantages of staining

microorganisms.
 Materials

Clean glass slide, sterile toothpick,

microscope, immersion oil, xylol,
cotton, methylene blue staining
solution
 PROCEDURES:
1. Clean glass slide.
2. Place slide on the desk; with a wire loop, put a small drop of tap water on the center of the slide.
3. With a sterile toothpick, obtain material from around the margins of your gums or between the teeth. Mix it into the
drop of water on the slide and spread out to make a thin film. Allow the slide to air dry on level surface. Drying may
be hastened by holding the slide film side up over a gentle flame.
4. Fix smear by passing he slide through the flame, film slide up, three times. The slide should not be so hot that it can
not be tolerated when touched to the back of your hand. Allow to cool.
5. Place the slide on wire staining rack in the sink and cover smear with methylene blue
solution. Allow stain to act for 1 minute.
6.Pour off stain and wash slide gently with tap water to remove excess methylene blue.
7.Shake off water and blot carefully (do not rub) with filter paper.
8.Examine under high power objective and later on the oil immersion objective.
9. Make labeled sketches of representative morphological types seen.
 GROUP 5 DRAWING APRIL 26, 2021

Bacteria: Staphylococcus epidermidis Bacillus megaterium

Total Magnification: 1000x 1000x
Morphology (shape): cocci/grapes rods
Arrangement of cells to
clusters rods
one another:
Color: appeared purple in methylene appeared pink in methylene
blue blue
 • Of what value is a simple stain?

Simple staining can be used for all types of bacterial cells to give

contrast to the other- wise colorless cell in order to determine cell
morphology, size, and cell grouping. This technique
is simple because only one dye is used and direct because the
actual cell is stained.
 2. Can dyes other than methylene blue be used
for direct staining?
Yes, due to the negative charged cell wall, bacteria can absorb the
positively charged stains such crystal violet, basic fuchsin, and
sufranin.
 3. What would happen if no heat fixing was
done?

If there is no heat fixing done the bacteria will not stick or adhere
to the glass slide, that will cause the bacteria to be wash off easily
or leaving to none to be seen under the microscope.
 4. Do all organisms on your slide appeared to be
identical?

No, they are radically different because some microorganism has

its own shape and size, and the types of dyes it absorbed. By
looking on its characteristic we can identify the microorganisms,
and they differ not only in size but also in structure and other
characteristics.
 5. What conclusion can you draw from your
findings?

The conclusion that we can draw from the findings is that with the
help of staining, it helps to know the different morphological types
of bacteria since it increases the visualization of the bacteria under
a microscope, and also considering the procedures of staining
would lead to the desired outcome.
 6. List at least three types of bacteria whose names reflect their
shapes and arrangements, and state the meaning of each name.

First is the Streptococcus (coccus shape), it is spherical and this is

a strep throat. Next is the Lactobacillus (Bacillus shape), it is rod
shape found in probiotics. Lastly, the Spirillum Pulli ( spirillum
shape), it is spiral shaped.

Streptococcus Lactobacillus Spirillum Pulli

GORUP 5 APRIL 26, 2021

Th an k Have a
great day
ahead.

yo u!