Human Red Blood Cell and White Blood Cell

Counting using Hemocytometer

Allan R. Dupla1
1Biology Student, Department of Biology, College of Science, Polytechnic
University of the Philippines

ABSTRACT
Blood is important in the human body, they function in delivering oxygen and
nutrients and the removal of carbon dioxide and other waste product in the body.
Hemocytometer was used in counting blood cells and many types of microscopic
substances. The purpose of the study to know the range of the human red blood cell
(RBC) and white blood cell (WBC). The result show that RBC is always greater than
WBC, the expected value of RBC is within 4,000,000/L to 5,500,000/L and a WBC
of 5,500/L to 10,000/L, while the experiment show that the average RBC is
2,598,000/L and 300 WBC it tell us that there is a deficiency of the human blood
RBC and WBC.
Keyword: Blood, Hemocytometer, Red Blood Cell, White Blood Cell

INTRODUCTION
Blood circulates through the
body bringing O2 and nutrients to the
tissues and removing CO2 and other
waste products. As it moves around
the body it aids interchange between
the fluid compartments, dissipates
heat and distributes hormones, thus
helping to maintain homeostasis and
to coordinate the activities of the
various organs. In addition blood
contains haemostatic components that
control bleeding. Finally, it performs a
role in defending the body against
foreign invaders as it carries cells and
antibodies that seek out and destroy
microorganisms and foreign proteins.
Blood can be separated into two
components - a yellowish fluid,

plasma,
and
cells
which
are
suspended in it. Plasma is that part of
the extracellular fluid which is
restricted to the blood vessels. The
cells are of three kinds - red cells
(erythrocytes), white

cells
(leucocytes)
and
platelets
(thrombocytes) (Anand et al., 2012).
The hemocytometer is a device
originally used to count blood cells (as
the name suggests). It is now used to
count other cells and many types of
microscopic particles. It consists of a
thick glass microscope slide with a
rectangular indentation that creates a
chamber of certain dimensions. This

chamber is etched with a grid of

perpendicular lines (Piuri and Scotti,
2004).

The formula below was used for the

computation of number of red blood
cells.

The study aims to know the

number of Red Blood Cell (RBC) and
White Blood Cell (WBC) of the human
blood using hemocytometer and to
know the significance of blood
counting.

METHODOLOGY

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

(10)
Where:
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

RBC Counting

WBC Counting

In this
experiment human
blood was used, first aspirate 5 L of
blood sample then dilute the blood
sample by adding 995 L of isotonic
saline solution after that place the
solution on a microcentrifuge tube,
homogenize the solution by shaking it
side by side about 5 times. Carefully
transfer the blood solution to the
hemocytometer let the drop come in
contact with the ruled area near the
edge of the cover slip so the fluid will
flow under the cover slip by capillarity
then place the counting chamber in
the stage of the microscope and using
LPO focus on the central big square
(millimeter) of the counting chamber.
Count the red cells in 5 medium
squares preferably those on the
corners and one somewhere in the
middle. Each medium square contains
16 small squares. To avoid counting a
cell twice, RBC touching the upper and
left borders of a given square are
included while those touching the right
and lower borders are not included.

The steps in WBC counting are almost

the same in RBC counting.
But the formula used is different,
below was used for the computation of
number of white blood cells.
Number of cells/mm3 = (L) (20) (10)
Where:
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
RESULT AND DISCUSSION
Figure 1. The result for white blood
cell counting

WBC Count
350
300
250
200
WBC COUNT 150
100
50
0

TRIALS

Figure 2.The result for red blood cell

counting

RBC Count
2900000
2800000
2700000
2600000
2500000
RBC Count
2400000
2300000
2200000
2100000
1

TRIALS

Table 1. The result for the expected

and observed value for RBC and WBC.

Observ
ed
Value
(Ave.)
Expecte
d Value

RBCs/L

WBCs/L

2,598,000

300

4,000,0005,500,000

500010000

As shown in the figure 1 and 2 the

range
of
RBC
observed
was
2,395,000/L to 2,790,000/L in the
average of 2,598,000/L while the
WBC ranging within 250/L to 325/L
in the average of 300/L as seen in
table 1, the normal value of the
human
RBCs
range
within
4,000,000/L to 5,500,000/L and a
WBC of 5,500/L to 10,000/L. Theres
a big difference in the experiment
observation unlike in the expected
frequency of RBC and WBC. RBC is
always greater than WBC.
The significance of irregularity in
count, an increase in the WBC count in
blood signifies the presence of an
infection, since the body produces
more WBCs as it fights against the
infection. A low WBC count may
decrease the body's ability to fight
disease-causing germs like bacteria
and viruses. Anemia is a decrease in
normal number RBCs or less than the
normal quantity of hemoglobin in the
blood. In the various types of anemia,
iron deficiency anemia is the most
common. It occurs when the dietary
intake or absorption of iron is
insufficient, and hemoglobin (which
contains iron) cannot be formed
(Abdul et al., 2009). So the result tells
us that the patient maybe undergoing
a blood disease because of the lack of
RBC and WBC but maybe there is a
wrong procedure that can affect the
result of the blood counting.
CONCLUSION
RECOMMENDATION

AND

Based
on
the
results
of
the
observation, numbers of WBCs are

always lesser than the number of

RBCs. The normal range of human RBC
is within 4,000,000/L to 5,500,000/L
and a WBC of 5,500/L to 10,000/L.
The result of observation tells that
maybe the person is undergoing a
disease cause by lack of human blood
RBC and WBC or maybe theres a
wrong procedure that affect the blood
counting. For recommendation, to
avoid the error in blood counting make
sure that the step by step process is
well done and also better to have
more trials.
REFERENCES
Abdul Nasir, A. S., Mustafa, N., Mohd
Nasir, N.
F.,
Application
Of
Thresholding
Technique
In
Determining Ratio Of
Blood Cells
For Leukemia Detection, Proceedings
Of The International
Conference

On Man-Machine Systems,
13 October 2009.

11

A. Anand, V. K. Chhaniwal, N. R. Patel,

B.
Javidi, Automatic Identification
Of
Malaria-Infected
Rbc
With
Digital
Holographic Microscopy
Using Correlation Algorithms, IEEE
Photonics
Journal,
Volume
4,
Number 5, October
2012.
Vincenzo
Piuri,
Fabio
Scotti,
Morphological
Classification
of
Blood Leucocytes By
Microscope
Images,
IEEE
International
Conference On Computational
Intelligence For Measurement
Systems
And Applications, Boston,
Md, Usa, 14- 16 July 2004.

APPENDIX

Table 2. The raw data for red blood cell counting.

TRIAL

Chamb
er

tota
l

1 (1&2)

C1

57

51

49

55

54

266

C2

46

50

57

53

51

257

C1

52

61

58

46

33

250

C2

42

51

40

44

52

229

C1

61

56

55

52

48

272

C2

59

62

60

52

52

285

2 (3&4)

3 (5&6)

4 (7&8)
5
(9&10)

C1

216

C2

265

C1

51

63

54

60

55

283

C2

55

65

49

58

48

275

Densit
y
266000
0
257000
0
250000
0
229000
0
272000
0
285000
0
216000
0
265000
0
283000
0
275000
0
Averag
e

Table 3. The raw data for white blood cell counting

TRIAL
1
2
3
4
5

Cham
ber
C1
C2
C1
C2
C1
C2
C1
C2
C1
C2

A
2
1
2
3
2
2
3
2
2
0

B
3
2
2
1
1
2
1
2
2
3

C
2
0
0
0
1
0
0
1
1
1

D
0
3
1
1
3
2
1
1
3
1

tot
al
7
6
5
5
7
6
5
6
8
5

Densit
y
350
300
250
250
350
300
250
300
400
250
Avera
ge

Avera
ge
325
250
325
275
325

300

Avera
ge
261500
0
239500
0
278500
0
240500
0
279000
0
25980
00