Lab 5 Aniphy Blood Count PDF

Evaluation of the Blood Cell Count
using Hemocytometer
Aduna, Naomi Bless S.
Department of Biology, College of Science, Polytechnic University of the Philippines
ABSTRACT
Blood counting looks for abnormalities in the blood, such as unusually high or low numbers of
blood cells. It is a common test that can help to diagnose a wide range of illnesses, infections and
diseases. The use of hemocytometer has long been used and is one of the earliest device used
in blood counting. The aim of the study is to determine the levels of RBC and WBC in the blood
samples obtained from human body through the hemocytometer. Results showed that the RBC
total count obtained was 2,660,000 cells/mm3 which is below the normal range of 4.5 million for
males and 4 million for females per mm3. On the other hand, low levels of WBC was also observed
which was only 555 cells/mm3. Normal range is from 4,500 to 10,000 cells/mm3. The levels may
indicate abnormalities coupled with high risk of developing different diseases such as heart failure,
renal infection, cancers, etc. Abnormal increase or decrease in cell counts as revealed in a
complete blood count may indicate underlying medical condition
Keywords: blood count, hemocytometer, RBC, WBC, diagnosis
INTRODUCTION
A complete blood count is a
measurement of the number of blood cells an
individual has in circulation based on laboratory
evaluation of a blood sample. A viable cell count
is crucial for the study of eukaryotic cells for
different purposes such as the management of
cell cultures in biological research, and the
titration of cell populations in diagnostics
(Herrera et al., 2015). A complete blood count
test measures several components and features
of the blood, including red blood cells, which
carry oxygen and white blood cells, which fight
infection. (Chabot-Richards and George 2014).
Red blood cell (RBC) concentration and white
blood cell (WBC) concentration are the major
parameters corresponding to a specific disease
or disorder.
The complete blood count (CBC) is

traditionally performed with a hemocytometer, a
device designed for cell counting comprising a
microscopic glass slide with a rectangular
indentation that creates a chamber 100 m
deep, under a conventional optical microscope.
(Roy et al., 2014). The hemocytometer is the
most widely used device for determining cell
concentrations, requiring consistent criteria and
tenacity to obtain measurements correctly and
reproducibly. (Stone et al., 2009)
The aim of the study is to evaluate the
blood count of a blood sample from a human
using a hemocytometer, specifically the RBC
and WBC total count present in circulation.
METHODOLOGY
RBC Counting
Human blood sample was obtained by
pricking and was placed in a container. 5L of
the blood sample was diluted by adding 995 L
of isotonic saline solution. The solution was then
transferred to a microcentrifuge tube and has
been shaken side by side to homogenize the
solution. A small amount of blood solution was
transferred to the hemocytometer. The counting
chamber was placed under the microscope. The
microscope was focused on the central big
square (mm) of the counting chamber using
LPO. Red cells were counted in the 5 medium
squares. Each medium square contains 16 small
squares. To avoid counting a cell twice, RBC
touching the left and upper borders of a given
square were included in the count, while those
touching the right and lower borders were not
included in the count of that given square. The
complete blood count was computed using the
following formula:
acetic acid to permit the rupture of RBC. The

solution
was
then
transferred
to
a
microcentrifuge tube and has been shaken side
by side to homogenize the solution. A small
amount of blood solution was transferred to the
hemocytometer. The counting chamber was
placed under the microscope. The microscope
was focused on the central big square (mm) of
the counting chamber using LPO. White cells
were counted in the 5 medium squares. Each
medium square contains 16 small squares. To
avoid counting a cell twice, RBC touching the left
and upper borders of a given square were
included in the count, while those touching the
right and lower borders were not included in the
count of that given square. The complete blood
count was computed using the following formula:
Number of cells/mm3 = ()(20)(10)
L = average number of WBC in 1 big square (Total

number of WBC in 4 big squares / 4)
20 dilution factor
10 factor of depth
Number of cells/mm3 = (80)(400)(200)(10)
E = number of RBC counted in the 5 medium (80

smalls) squares
400 total number of small red square
200 dilution factor
10 factor of depth
WBC Counting
RESULTS AND DISCUSSION

The activity was planned to determine
the total blood count (RBC and WBC) from blood
samples that were obtained from human body
through the use of hemocytometer. The total
count obtained from the 5 media and standard
error are summarized in Table 1 and 2. Table 1
shows the RBC total count of 2666000
cells/mm3 that is lower than the normal range
Human blood sample was obtained by

pricking and was placed in a container. 5L of
the blood sample was diluted by adding 1%
No. of Cells/mm3
NO. of Cells/mm3
which is 4.5 million RBC/mm3 for males and 4

million RBC/mm3 for females. Levels of RBC
observed were too low which indicates blood
abnormalities such as anemia. In table 2, the
WBC count are also presented. The total WBC
observed under the hemocytometer is 555
cells/mm3, far lower than normal which is 4,50010,000 cells/mm3, indicating that the levels of
WBC counted may also present abnormalities
coupled. e.g. Leukopenia (a reduction in the
number of white cells in the blood).
3000000
2000000
1000000
0
RBC
WBC
Blood Sample
Figure 1. Graph showing the levels of RBC vs WBC
observed using the hemocytometer under the microscope
(LPO).
Ave
Density per
mm3
58
50
46
53
53
52
43
54
89
62
50
47
48
77
52
62
44
44
55
58
51
38
41
60
46
55
44
47
67
54
546000
444000
466000
668000
542000
TRIAL
Table 1. Total RBC count (cells/mm3)
1
2
3
4
5
2666000
S.EE
3.93
Ave
Density
per mm3
100
3.5
175
1.5
75
2.5
125
1.6
80
TRIAL
Table 2. WBC count (cells/mm3)
555
S.E
18.26
It is clear from the results that if an

individual possesses the observed blood count,
diseases coupled with the abnormalities
observed in the blood samples may be present.
Blood disorders affect one or more parts of the
blood and prevent the blood from doing its job.
They can be acute or chronic. Many blood
disorders are inherited. Other causes include
other diseases, side effects of medicines, and a
lack of certain nutrients in your diet. (Madjid
2013).
Red Blood Cells are the major cellular
component of human blood which are
responsible for gaseous exchange between
living cells and external environment. (Rakshit
and Bhowmik 2013). Blood cell count is one of
the earliest parameters established to
quantitatively study the blood and aid the
diagnosis (Verso, 1964). Red blood cell (RBC)
volume variation has recently been shown to
predict a wide range of mortality and morbidity.
Death due to cardiovascular disease, cancer,
infection, renal disease, complications in heart
failure and coronary artery disease, advanced
stage and worse prognosis in many cancers,
poor outcomes in autoimmune disease, and
many more. (Patel et al., 2015). The levels of
RBC observed in the study is at risk of
developing such diseases.
3
On the other hand, White blood cell count

is also an essential aspect of the complete blood
count. White blood cells act in response to
infections or injuries, and play an important role
in our body's defense system. Most white blood
cells migrate from the circulating system to the
tissues in order to soothe local conditions. Other
white blood cells trigger adaptive immune
system to eliminate pathogens and provide longlasting effect. Such responses would lead to
abnormal circulating white blood cell count.
(Tefferi et al., 2005). The levels of WBC
observed in the study were too low and may also
have high risk of developing low WBC related
diseases. A lower count of WBC, a condition
known as leukopenia, is often associated to
bone marrow deficiency, certain viral infection,
and severe bacterial infection (Carli et al., 2015).
Although the method of blood counting

through the use of hemocytometer is more
easier compared to other procedures, some
disadvantages of using such technique are also
present. For example, dead cells that are not
identified from lives which were included in the
count, small cells that are difficult to locate and
even impossible to mention, and precision is also
tough to achieve. The technique is also quite
tedious for it requires patience and longer time in
counting individual cells in the chambers of the
hemocytometer.
CONCLUSION
A complete blood count is a blood test
used to evaluate overall health and detect a wide
range of disorders, including anemia, infection
and leukemia. Abnormal increase or decrease in
cell counts as revealed in a complete blood
count may indicate underlying medical condition.
LITERATURES CITED
Carli, L , Tani C , Vagnani S , Signorini V , Mosca
M.
2015. Leukopenia, lymphopenia
and
neutropenia in
systemic
lupus erythematosus: Prevalence and
clinical
impact. A systematic
literature review,
Seminars
in
Arthritis and Rheumatism
Chabot-Richards, Devon S. MD., Tracy I.
George, MD. 2014. White Blood Cell
Counts Reference
Methodology . Clin
LabMed(2014)://dx.doi.org/10.1016/j.cll.
2014.10.007
Herrera, Daniela Cadena, Joshua E. EsparzaDe Lara,
Nancy
D.
RamrezIbaez, Carlos A. LpezMorales,
Nstor O. Prez , Luis F.
FloresOrtiz, Emilio Medina-Rivero. 2015.
Validation of three viable-cell counting
methods:
semi-automated,
and
automated.
BiotechnologyReports
Madjid M, Fatemi O. 2013. Components of the
complete blood count as risk predictors
for
coronary heart
disease.
Tex Heart Inst J
2013;40:17e29.
Patel, Harsh H., Hasmukh R. Patel and John M.
Higgins. 2015. Modulation of red
blood cell
population
dynamics
is a fundamental
homeostatic
response
to
disease.
American
Journal of Hematology,
Vol.
90,
No. 5, May
2015
Rakshit, Pranati and Kriti Bhowmik . 2013.
Detection
of Abnormal Findings in
Human RBC in
Diagnosing
Sickle Cell Anemia Using
Image
Processing. Procedia Technology 10
pp. 28 36
Roy, M, G. Jin, D. Seo, M.-H. Nam, S. Seo. 2014.
A simple and low-cost device performing
blood cell
counting
based
on
lensfree shadow imaging technique,
Sensors
and Actuators
B:Chemical
Stone, Laurel Rebecca, Diane R Gray, Krissy

Remple and Matthew P. Beaudet. 2009.
Accuracy and precision comparison of
the hemocytometer and automated cell
counting methods. Faseb
Journals
Verso, M. L. 1964. "The Evolution of BloodCountingTechniques". Med.Hist. 8 (2): 1
49158.

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Evaluation of the Blood Cell Count

Keywords: blood count, hemocytometer, RBC, WBC, diagnosis

The complete blood count (CBC) is

acetic acid to permit the rupture of RBC. The

Number of cells/mm3 = ()(20)(10)

L = average number of WBC in 1 big square (Total

Number of cells/mm3 = (80)(400)(200)(10)

E = number of RBC counted in the 5 medium (80

RESULTS AND DISCUSSION

Human blood sample was obtained by

which is 4.5 million RBC/mm3 for males and 4

Table 1. Total RBC count (cells/mm3)

Table 2. WBC count (cells/mm3)

It is clear from the results that if an

On the other hand, White blood cell count

Although the method of blood counting

Stone, Laurel Rebecca, Diane R Gray, Krissy

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