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NUMBER 8 COUNT
ARUGAY BAYONGASAN CAUILAN JOSE LIZARDO PAGADDU
TAGUINOD
INTRODUCTION
White blood cells or leukocytes -- spherical, motile, and
nucleated cells of varying sizes
The cover slip was placed over the chamber and the
chamber was placed on the microscope stage, and
the pipette was rolled again before charging. The
first 2-3drops were discarded.
It may be used to determine presence of other diseases that affect WBCs such
as allergies, leukemia or immune disorders. It is also done as a follow up test
on patients on chemotherapy or to monitor bone marrow function.
2. What other tests has to be done with total count to make
it more meaningful?
CBC includes all kinds of cell count especially with RBCs, WBCs and
platelets. Along with total leukocyte count, other tests come with to
make sense of some cases.
The closest one is called the differential leukocyte count (DLC) which
accounts for the individual amount of major types of WBC. Tests like
RBC, PCV, Hgb, red blood cell indices, thrombocyte, and mean platelet
volume count may be added with total leukocyte count as components
of a complete blood count (CBC).
3. Why should you destroy the RBCs while doing WBC count?
The RBCs are hemolyzed by a glacial acetic acid present in Turks fluid while
it does not affect WBCs. A dye stains the nuclei of leukocytes to be able to
recognize the minute WBCs.
4. What is the normal range of total WBC count? List three
important physiological variations?
- Physiological variations can also be called as diurnal variations that affect WBC count:
1. A pregnant woman in labor at the last month of pregnancy may have increased WBC levels.
2. Smoking may also cause increased WBC count.
3. Newborns with normal health status have higher WBC count than adults.
4. Cases such as morning pseudoneutropenia wherein WBC count tends to be lower in the
morning and higher in the late afternoon. This is noticed in some patients who are taking
antipsychotic medications.
EXPERIMENT DIFFERENTIAL COUNT
NUMBER 9 OF WBCs
ARUGAY BAYONGASAN CAUILAN JOSE LIZARDO PAGADDU
TAGUINOD
INTRODUCTION
Differential Count of RBCs - white blood cell types are
separately counted from each other manually in the Neubauer
chamber
Neutrophils - 50-70%
Eosinophils - 1-4%
Basophils - 0-1%
Lymphocytes - 20-40%
Monocytes - 2-8%
APPARATUS
Microscope
Clean dry grease free glass slides with even edges
Cedar wood oil
Staining rack
Lancet, cotton and spirit
A drop battle containing distilled water
A drop battle containing Leishmans stain
Eosin
Methylene blue
Acetone-free methyl alcohol
PROCEDURE
Preparation of blood film
1. The blood smear was placed on the staining rack then Leishmans stain
was added drop by drop until the entire film was covered. The number of
drops added were counted.
2. The undiluted stain was allowed to act for 2 minutes but wasnt allowed to
dry up. The cells were fixed during these 2 minutes.
3. Double the number of distilled water drops were added to dilute the stain
and then it was mixed gently.
4. The stain was not allowed to dry and after 7
minutes the stain was drained off.
5. The slide was washed in running tap water until
the film turned pink and was kept in vertical
position to drain and dry.
The slide is slowly moved towards the tail end and the
cells are counted. The slides is then shifted up and
moved in opposite direction. This pattern of movement
of slides (zigzag pattern) takes into consideration all
the parts of the film and ensures that a cell is not
counted more than once.
Neutrophil 50-70 % 44 %
Eosinophil 1-4 % 4%
Basophils 0-1 % 0%
Agranulocytes
Lymphocytes 20-40 % 51 %
Monocyte 2-8 % 1 %
The experiment was done by identifying each white blood cell seen and
was recorded until 100 WBCs were counted.
When reporting a manual differential count, the results are reported as
the percentage of each cell type present as seen in the table above.
The patient had a slightly decreased Neutrophil count and a slight
increase in Lymphocyte count.
These results can be correlated to the condition of the patient, since she
is currently having cough and colds, the neutrophils will then slightly
decrease and the lymphocytes will increase due to viral infections. The other
cell types had a normal result.
Other factors can be considered, like for the stain used, the
performers increased the contact time between the stain and the smear.
Since the stain used seems to be contaminated or old, this also gave a
hard time for the performers to identify the cells and they also counted
not only in the thin portion near the feathery edge, but continued to
count in the thick portion of the smear. Nevertheless, they were able to
count 100 WBCs.
ANSWERS TO QUESTIONS
1. What is the purpose of methyl alcohol in Leishmans stain?
5. What are the terms which refer to (a) increase and (b) decrease in the
lymphocyte count?
Lymphocytosis Increase in the number of lymphocyte
Lymphocytopenia Decrease in the number of the lymphocytes
6. Name two conditions in which each occur
a. Lymphocytosis
i. Chronic Infection i.e. TB, Syphilis and
malaria
ii.Viral Infection
b. Lymphocytopenia
i. Acute infection and illnesses
ii.Hodgkins disease
7. What is Monocytosis? When does it occur?
12. What are the special features of monocyte? How will you
differentiate monocyte from large lymphocyte?
a. Monocyte
i. Nucleus Large, centrally located and kidney-shaped
ii.Cytoplasm Large amount of pale grayish-blue cytoplasm and
no granules seen
Monocyte versus Large Lymphocyte
i. Size
1.Lymphocyte - cell is twice as big as the Red Cells
2.Monocyte - cells is about 2.1/2 to 3 times
ii.Nucleus
1.Lymphocyte - completely fills the cells
2.Monocyte - big cell with a kidney shaped nucleus
iii.Cytoplasm
1.Lymphocyte - a bigger cell with a rim of cytoplasm all around
2.Monocyte - a big cell with a large amount of clear cytoplasm in relation to the
nucleus
13. How do you differentiate a bi-lobed eosinophil from a neutrophil?
They are differentiated through their granules
i. Neutrophil Fine neutral granules of Light Violet
color
ii.Eosinophil Coarse and Orange or Red color
14. What are the microscopic adjustment you will do before starting the
experiment?
a. The microscope should be adjusted for oil immersion lens
b. The condenser is raised
c. The diaphragm is completely opened and plane mirror is used
d. The oil immersion lens is made to touch the oil by viewing from the side.
e. The fine adjustment screw is adjusted till the cell are brought into focus.