HEMATOLOGY II

By:
Name : Vio Indah Budiarti
Student ID : B1B015009
Group : VIII
Sub Group :2
Assistant : Afrizal Vikri Avani

PRACTICAL REPORT OF ANIMAL PHYSIOLOGY I

MINISTRY OF RESEARCH, TECHNOLOGY, AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
BIOLOGY FACULTY
PURWOKERTO
2016
 I. INTRODUCTION

1.1 Background

Blood is the liquid found in all living organisms (except plants) that serves
to send a high-level substances and oxygen needed by the body tissue, transporting
chemicals products of metabolism, as well as the body's defense against viruses or
bacteria. The main function of blood is to carry oxygen from the lungs or gills to
body tissues. Blood has a composition comprising about 55% of blood fluid
(plasma) and 45% of blood cells. Blood-forming element includes three kinds of
blood cells, namely red blood cells (erythrocytes), white blood cells (WBCs), and
platelets (thrombocytes). Third blood cells are classified into solid elements called
corpusculer. Blood plasma is a yellowish clear liquid that element substantially
similar to the cytoplasm. Plasma consists of 92% water and contains a complex
mixture of organic and inorganic substances (Pearce, 2002).
Blood contains hemoglobin which serves as a binder oxygen. Most
invertebrates or invertebrates are small, oxygen is directly absorbed into the blood
plasma as a carrier protein dissolved oxygen freely. Blood is red because of their red
blood cells. Consists of red blood cells, white and platelets (Pearce, 2002).
The red blood cells are round flattened second concave surface. Red blood
cells have no nucleus and contain hemoglobin. White blood cells are not actually
white, but clear. Known to distinguish white blood cells from the red blood cells
red. White blood cells are irregularly shaped or non-permanent. Unlike red blood
cells are always inside the blood vessels, white blood cells can get out of the blood
vessels. The ability to move freely is required of white blood cells in order to
perform its function to keep the body. White blood cells have a cell nucleus but
no color or no pigment. Platelet round or oval. The size of the blood platelets are
smaller than red blood cells. The main function of red blood cells (erythrocytes)
is transferring hemoglobin. Normal erythrocytes are round or slightly oval with a
diameter of 7-8 microns (normosit). Viewed from the side, looks like a disc or
erythrocyte biconcave central akromia with approximately - the diameter of
the cell (Colorful, 2009) The amount is less than 300,000 in each 1 mm3 of blood.
Platelet-lived, only 8 days. Platelet function in the blood clotting process. When
injury occurs, blood out through the wound. Platelet touched the surface of the
wound, then broke and tissue factor (Pearce, 2002).

1.2 Purpose

The purpose of the practical is to understand the response of the red blood cells
against a wide range of media that has a concentration of osmotic different, can
determine the concentration of internal red blood cells, to understand the shape and
structure of the cells, comparing the shape and structure of the blood cells of frogs and
humans, and can understand the process blood clots and determine the length of time
the blood clotting in humans.
 II. MATERIAL AND METHOD

2.1 Material

The tools used in this lab is cotton, lancet, scissors, glass capillary vessels, glass
objects and glass cover, spuilt microscope, tweezers, pipette and stopwatch.
The materials used in this lab is a solution of NaCl 0.2%, 0.4%, 0.6%, 0.9%, 1.0%,
blood frog (Fejervarya cancrivora), and blood of humans (Homo sapiens), alcohol
70%, and anticoagulants: Na-citrate / EDTA

2.2. Method

Method that used in this practical are:

2.2.1 Concentration Blood Cells
a. Materials and equipment to be used is prepared.
b. Fingertips moistened with alcohol using a cotton swab, then prick your finger
with a sterile lancet, sequence toward the fingertip so that the blood deposited
in the cup which had previously been moistened with EDTA.
c. Blood on the cup dripped on glass objects with the aid of a pipette, then add a
drop of 0.4% NaCl solution, both homogenized by way shake.
d. The liquid mixture is immediately covered with a glass lid. If not promptly
closed evaporation will happen to change the concentration of NaCl solution.
e. The mixture was observed under a microscope.
f. Do the work steps above for the next blood droplets, using 0.2% NaCl, 0.6%
and 1.0%. Each mixture of blood at a certain concentration should be observed
under the microscope.
g. NaCl concentration determined in which red blood cells did not change shape.
2.2.2 Structure of Red Blood Cells
a. The preparation of human blood obtained by piercing the fingertip with a sterile
lancet that has previously been moistened with 70% alcohol, then the blood
that comes out can be immediately used for the experiment. In the object glass
clean and dry, blood dripped from his fingertips and then added a drop of 0.6%
NaCl solution for frogs and NaCl 0.9% for men.
b. Once the two are mixed and then covered with a glass cover and observed
under a microscope.
 c. The difference between these two cells were observed and made drawings of
each cell earlier.
d. Finger puncture marks had been cleaned with alcohol swabs, cotton can be kept
under pressure to the wound can be immediately shut down by the formation
of blood clots.
2.2.3 Time Frozen Blood
a. Finger cleaned with 70% alcohol, after the alcohol dries finger pricked with a
lancet sterile or disposable lancet.
b. Capillary tube affixed to a droplet of blood from a finger.
c. Interval with a time of 1 minute glass capillary vessels cut little by little until
it looks fibrin that forms marked with pieces of capillaries that remain attached
or hanging after the break.
d. Time takes blood to clot is recorded, namely the time since the finger hacked
to broken capillaries remain hanging.
3.2 Discussion

In hematology 2 practical class using human blood and frogs as preparations

observations. Human blood and frogs are used to determine blood cell response when
administered NaCl with different concentrations, determine the shape and sstruktur of
both these blood cells and find out the process and the length of time required in the
human blood clotting. Observations lab is divided per group where the group first
observed diameter and structure of blood cells before and after treated with a solution
of NaCl 0.2%, Group 2 observed diameter and structure of blood cells before and after
treated with a solution of NaCl 0.4%, Group 3 observed diameter and structure of red
blood cells before and after given a solution of NaCl 0.6%, group 4 observed diameter
and structure of blood cells before and after treated with a solution of NaCl 0.9% and
5 groups observing the diameter and structure of blood cells before and after given a
1% NaCl solution.
Based on lab results showed that human blood with a concentration of 0,2%
has a size of red blood cells by an average of 7,5 m; 0,4% concentration of 8 m, a
concentration of 0,6% at 10 m, 0,9 concentarion % of 9,8 m, a concentration of
1.0% at 6.5 m diameter blood cells. While the blood frog with a concentration of
0.2% memilki blood cell diameter of 16,5 m, 0,4% amounting to 17,25 m, 0,6% at
18 m, 0,9% 16,25 m and a concentration of 1% of 13 .
According Wiguna (2009), red cells in the frog (Fejervarya cancrivora) has an
oval shape and has a size greater than human erythrocytes. Mature erythrocytes oval
or elliptical, flattened and have a nucleus. Red blood cells including erythrocytes
owned frog larger than any other vertebrate animal. With the nucleus in erythrocytes
of frogs, it can minimize space for hemoglobin for oxygen yag required by frogs are
not only bound by red blood cells in the lungs, but instead of oxygen diffuse through
their skin. According Colorful (2009) red cells of normal blood cells appear round with
a diameter of 7,5 m with a rim thickness. From the side looks erythrocytes shaped
like a disc with a second concave surface (biconcav disk). According to the
observations during the practical observations frog red blood cell structure according
to the literature, but there are differences in the size of the diameter of this could be
error count cells with sizes big ones so that the end result is greater diameter than the
mentioned.
 The results of the observation time of blood clotting from groups 2,3,4, and 5
showed that 14 minutes, 3 minutes, 9 minutes 26 seconds and 3 minutes. According
Pharmaspica (2011), the blood clotting time is the time it takes from when the blood
out to form fibrin threads in the process of blood clotting. People with hemophilia, a
blood clot difficult. If the patient suffered minor injuries, can lead to serious bleeding.
The period of the normal bleeding for 1-3 minutes, from the description of the results
of five groups according to the literature, while in group 2, 3, and 4 do not correspond
with the library because there are other factors that influence it, perhaps a lack of
vitamin K in the body, resulting in difficult blood clotting .
In response to blood vessel damage, then a series of complex chemical
reactions that occur in the blood and involves a lot of blood clotting factors. The end
result is the formation of a complex substance known collectively activated
prothrombin activator (Pharmaspica, 2011). Thromboplastin formed because of
damage to the platelets, as long as there is the calcium salt in the blood will change
prototrombin into thrombin, causing a blood clot (Pearce, 2002). Agglutination or
clumping of red blood cells may be influenced by a variety of substances, and can
occur in the bloodstream at various pathological circumstances. Agglutinin present in
the plasma few individuals can cause erythrocyte agglutination others. Agglutinin
became the basis of blood empatbagian (Leeson, 1990). Blood clots needed four
factors:
1). Calcium salts that are normally present in the blood.
2). Cells were wounded liberating tissue factor.
3). Thrombin formed from prothrombin when tissue factor.
4). Fibrin is formed from fibrinogen in addition to thrombin.
(Leeson, 1990).
The mechanism of blood clotting is a process autocatalitic and "self-limited"
in which the formation of thrombin that plays a fairly address the effects of
antithrombin circulating serine protease inhibitor and another, immediately converted
fibrinogen into fibrin in gel form (Ville, 1988). Platelets are touching the rough surface
will break and remove the enzyme tissue factor (thromboplastin). The process is as
follows; Platelets broke prothrombin thromboplastin Ca ions Thrombin - Vitamin K
- Fibrinogen Fibrin (Leeson, 1990). Thrombin inhibitors are of great interest as
candidates for anticoagulants because thrombin, which cleaves fibrinogen into fibrin
to form the clot, is at the apex of the clotting cascade. We used DNA thrombin binding
aptamers (TBA) to inhibit thrombin and consequently coagulation. We then used
complementary DNA as an antidote because it can reverse TBAs effect by base-
pairing with it (Figure 1a). Selective excitation of two different NRs to release TBA
and its antidote enables the pair to act as an on/off switch for coagulation (Helena,
2013)
Calibration is the process of verifying that an accuracy of measuring
instruments in accordance with the design. Calibration is usually done by comparing
a standard that is connected with national and international standards and certified
reference materials. Calibration, in general, is the process of adjusting the output or
indication of a measurement device to match the magnitude of the standards used
within specified accuracy. For example, the thermometer can be calibrated so that the
fault indication or correction can be determined and adjusted (via calibration
constants), so that the thermometer shows the actual temperature in Celsius at specific
points on the scale (Pattern 1971).
The liver synthesizes most of the clotting factors, so it plays an important role
in blood clotting. Liver disease interfere with this synthesis can lead to difficulty
clotting. Vitamin K is very important in the synthesis of prothrombin and other clotting
factors in the liver. This vitamin absorption from the gut relies on Garama bile
produced by the liver. If the bile duct is clogged (eg by gallstones), then the ability to
form a clot will be reduced (Wiguna, 2009).
Extrinsic clotting mechanism starts from external factors blood vessel itself.
Thromboplastin (membrane lipoproteins) are released by cells damaged tissue
activates prothrombin (plasma proteins) with the aid of calcium ions to form thrombin.
Thrombin converts fibrinogen to dissolve, into insoluble fibrin. Threads of fibrin to
form a clot or fibrin nets, which capture red blood cells, platelets and close the blood
flow through the damaged vein (Wiguna, 2009).
The intrinsic mechanism for blood clotting take place in a more simple way.
Each factor protein in a state of inactivity; if one is activated, it will activate the
enzymatic activity further factor in the circuit, so there will be a series of reactions
(cascade of reaction) to form clots, once formed, will clot occupational contractile
proteins in platelets. Along with clot retraction, a fluid called serum out of the clot.
Serum is blood plasma without fibrinogen and with no other factors involved in the
clotting mechanism (Wiguna, 2009).
 IV. CONCLUSION

Based on practical work that has been done can be concluded as follows:
1. Red blood cells in the frog (Fejervarya cancrivora) has an oval shape and has a size
greater than human erythrocytes and nuclei.
2. The concentration of blood cells on a hypertonic environment will cause Crenation
(shrinkage) in the blood cells because the water inside the cells out into the
environment. While the blood cells in a hypotonic environment caused plasmolysis
in the blood cells due to water from the environment into the blood cells.
3. The mechanism of blood clotting is as follows; Platelets broke prothrombin
thromboplastin Ca ions Thrombin - Vitamin K - Fibrinogen Fibrin.
4. Time of blood clotting is the time it takes from when the blood out to form fibrin
threads in the process of blood clotting. Frozen blood of normal time 1-3 minutes.
While the results of the observation time of frozen blood from groups 2,3,4 and 5
didapakan results of 14 minutes, 3 minutes, 9 minutes 26 seconds and 3 minutes.
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