DIFFERENTIAL LEUKOCYTE

By :
Name : Devina Alifah
Student ID : B1B017010
Group : VII
Subgroup :2
Assistant : Siti Masrifah

PRACTICAL REPORT OF ANIMAL PHYSIOLOGY I

MINISTRY OF RESEARCH, TECHNOLOGY AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
FACULTY OF BIOLOGY
PURWOKERTO
2018
 I. INTRODUCTION

A. Background

Blood is the body's tissues are different from other body tissues, are in a liquid
consistency, circulating in a closed system is called as blood vessels and functioning
transport various materials as well as homeostatic functions (Legler, 1997). According
to Banerjee et al (2017), blood components are consisted of four parts red blood cells
(erythrocytes), white blood cells (leukocytes), platelets, and plasma.
According to Nusa et al (2015), leukocyte types consist of 5 types, including:
neutrophils, eosinophils, basophils, monocytes and lymphocytes. Leukocyte play a role
in defending the body against ingress of foreign bodies is always deemed to have the
possibility to bring danger to the survival of the individual in addition or also called
have a role as immune system, white blood cells to function as a carrier of fatty
substances. Leukocytes are amoeboid or do not have a fixed shape (Hoffbrand, 1987).
According to Sutardi (2016), the immune system is a system in the body that
consisted of cells which produce certain compounds and these compounds work
collectively and are coordinated to fight foreign objects such as germs or poisons that
enter the body. The mechanism involves a combination of cells, molecules, and tissues
that play a role in resistance to infection that caused by various pathogenic elements
which found in our environment such as viruses, bacteria, fungus, protozoa and
parasites. And also other ingredients for microbes are called immune responses
(Baratawidjaja, 2000). White blood cell differential counting is one of the most
frequently performed blood tests for diagnosing diseases such as anemia, leukemia, and
HIV. Whereas, it needs time-consuming to count white blood cell types manually. In
addition, the result of diagnosis is subjective and depends on the expertise of the doctor.
An automatic white blood cell differential counting system is therefore preferred
(Fatichah et al, 2015).

B. Purpose

The objectives of this laboratory activity is to know the type of leukocyte and its
shape.
 II. MATERIAL AND METHODS

A. Material

The materials that used in this practice are methanol absolute 96%, alcohol 70%,
immersion oil, water, giemza 7%, tissue, and blood of human.
The tools that used in this practice are microscope, object glass, cover glass,
lancet, beaker glass, and drop pipette.

B. Methods

1. The object glass is cleaned by alcohol 70%.

2. The blood sample is taken using lancet and drop it on the object glass.
3. The blood sample is smeared using another object glass, the edge in front of the the
sample is touch using another object glass with 45 degree angle until the blood is
capillarity and pushed toward quickly.
4. The blood smear is fixed by methanol absolute for 5 minutes.
5. The blood smear is dry air.
6. The blood smear is soak by giemza 7% for 20 minutes.
7. The blood smear is rinse by the water.
8. Observed on the microscope.
 III. RESULT AND DISCUSSION

A. Result

Table 3.1. Observation Result of Differential Leukocyte in Human Entourage VII

Group Differential Leukocyte

Neutrophile Eosinophile Basophile Monocyte Limpocyte

1    - 

2   - - 

3  - - - 

4     

5   - - 

Figure3.1. Neutrophile Shape of Figure 3.2. Eosinophile Shape of

Leukocyte Magnification 400 X Leukocyte Magnification 400 X

Figure 3.3. Lymphocyte Shape Figure 3.4. Basophile Shape

Leukocyte Magnification 400 X Leukocyte Magnification 400 X
Figure 3.5 Monocyte Shape of
Leukocyte Magnification 400 X
B. Discussion

Based on the observation that we obtain, in our blood smear sample of leukocyte
only found neutrophile, eusinophile, and lymphocyte. While in the group 5 there are
found neutrophile, eusinphile, basophile, monocyte and lympocye. It is accoedance to
Nusa et al (2015) leukocyte types consist of 5 types, including: neutrophils, eosinophils,
basophils, monocytes and lymphocytes.
The stages in making blood smear preparations include blood sampling, blood
film making, drying, fixation, drying, coloring, washing, and labeling. Each stage has
different functions and purposes. Taking blood samples is intended to take proband
blood with the help of blood lancet pen, then making blood films to make blood smear
results. The blood smear must be as thin as possible so that it can be observed and the
blood cells do not accumulate and get closely. Drying is done with the help of the wind
so that the smeared blood quickly dries so that when done fixation does not fade.
Fixation aims to make the cell elements die but still maintain their shape, structure, and
size. The main function of fixation is to maintain the structure of blood cells that are
made into objects, change the refractive index of blood cells to be easily observed, and
change cells to easily absorb dyes. Drying is done so that the cells are perfectly fixed,
the remaining fixation evaporates and the smear results remain dry and do not fade
when stained. Staining using Giemsa which consists of methylene blue and eosin which
gives blue color to the cell nucleus. Then drying is done so that the color sticks perfectly
and washing is done so that the non-coloring dyes of the cells dissolve into the water
stream. Sterile distilled water is used so that there are no other microorganisms attached
to the blood smear because when an observation is made there can be an error of
analysis (Handari, 2003).
The formation of white blood cells starts from the early differentiation of
pluripotent hemopoietic stem cells into various cell types. In addition to these cells, to
form erythrocytes and form leukocytes. In leukocyte formation there are two types of
eelocytic and lymphocytic. Myelocytic leukocyte formation begins with young cells in
the form of myeloblasts whereas lymphocytic leukocyte formation begins with young
cells in the form of lymphoblasts. Leukocytes formed in the bone marrow, especially
granulocytes, are stored in the marrow until these cells are needed in the circulation.
Then, if the need increases, several factors such as cytokines will be released. Under
normal circumstances, granulocytes circulating in the whole blood are approximately
three times the amount stored in the marrow. This amount corresponds to the supply of
granulocytes for six days. While most lymphocytes will be stored in various lymphoid
areas except in a few lymphocytes which are temporarily transported in the blood
(Mohammad, 2001).
White blood cells have characteristics, among others, is colorless, has a nucleus,
loss of Hb, irregular shape, can move, and can be deformed. White blood cells can be
differentiated into two types, namely granulocytes and agranulosit. Actually, both types
of white blood cells is clearly visible on granulocytes. Granules contain a variety of
enzymes and proteins that help white blood cells to protect the body (Hoffbrand, 1987).
According to Arslan et al (2014), the shapes of white blood cells are more regular and
compact compared with red blood cells, which look like thick-edged rings. Moreover,
normal white blood cells are sparsely distributed across the background whereas
leukemic cells tend to grow over each other forming cell clusters. The shape
information is also useful to decompose these leukemic cell clusters into individual
cells.
Leukocytes are cells containing a nucleus, also called white blood cells. Seen in the
light microscope white blood cells have two types, namely, specific granules
(granulocytes) and agranulocyte. There are three types of granular leukocyte
neutrophilse is the largest group of white blood cells, makes up 45 to 75 percent of
white blood. Neutrophils are also referred to as polymorphonuclear (PMN), because the
nucleus has various types of forms and is segmented. Neutrophils are phagocytes, have
a role in fighting bacterial and virus infections; basophils represent less than 3 percent
of white blood cells. Basophils are 10-12 µm in diameter, with two core nuclei or
irregular core shapes. Basophils have a role in allergic reactions by releasing histamine,
which causes blood vessels to dilate; and asidofil (or eosinophils) are present 7% of
white blood cells and increased associated with the presence of asthma, allergies and
fever. Eosinophils have a diameter of 10 to 12 micrometers. Eosinophils are a group of
granulocytes that are responsible for fighting parasites which have a period of 8 to 12
days. Eosinophils contain profibrinolisin, thought to play a role in maintaining blood
from clotting (Effendi, 2003).
There are two types of leukocytes agranular are lymphocytes (small cells) are
agranulocyte leukocytes which have varying sizes and shapes. Lymphocytes are the
only type of leukocytes that do not have phagocytic abilities. Lymphocytes have the
main function of producing antibodies in response to foreign objects that are phagocytic
macrophages. Lymphocytes can be classified into two namely B lymphocytes and T
lymphocytes; monocytes (a rather large cell cytoplasm containing more) are the largest
leukocytes with a diameter of 15-20 µm and the population ranges from 3-9% of the
leukocyte total. Monocyte cytoplasm is pale gray blue and oval nucleus like a kidney or
horseshoe. The function of monocytes is to clean debris cells produced from the
inflammation or infection process, processing several antigens attached to the
lymphocyte cell membrane to become more antigenic so that they can be easily digested
by monocytes and macrophages, destroying foreign substances that enter the body
(Colville & Bassert, 2008).
 IV. CONCLUSION

Based on the observations it can be concluded there are two type of leukocyte
Granular (basophils, eosinophils and neutrophils ) and Agranural (monocytes and
lymphocytes).
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