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THE BLOOD
Introduction
Blood is a special tissue that supplies oxygen and nutrients to the tissue. The blood travels
through the blood vessels which is found all over the body of an organism. The main components
of blood are the plasma, erythrocytes, leucocytes and platelets. The plasma is identified as the
straw-colored component of the blood that carries cells and proteins throughout the body. The
blood cells that are produced in the bone marrow which contains the hemoglobin and supplies the
oxygen throughout the body are the erythrocytes. While the leucocytes are the white blood cells
which are the main defense of our body against foreign organisms that may enter our system.
The blood also has an antigenic property, this is due to the presence of antigens on the red
blood cell’s membrane. The human blood can be classified into grouping system based on the
presence or absence of the specific antigens, it can be classified as group A, B, AB and O, based
When injured the body has its own way of preventing much or more danger from
happening. Haemostasis is one of the mechanisms of the body that prevents it from losing blood
when injured. Prolonged bleeding and abnormal blood clotting may denote an interference in the
haemostasis process of the body. The clotting time of the blood must be not so slow to prevent
losing too blood that may be later on cause more serious damages to the injured person.
Materials and Method
Hemocytometer
square which is divided to nine 1 mm2 squares. The center square of the hemocytometer is also
divided into 25 smaller squares which is also subdivided into much smaller 16 1/25 mm2 squares
which are used for counting the red blood cells in the given blood samples.
To examine the hemocytometer and to proceed with the cell counting, it was done under
the microscope. The LPO was first used to locate the center square and then changed it to HPO to
focus on the much smaller squares. Cells touching or overlapping on the upper and left border of
the small squares was not counted, only the right and lower borders was included in the count.
Using the 200-cc distilled water the 0.5 grams mercuric chloride, 5 grams of sodium sulfate
and 1 gram of sodium chloride was dissolved. It was filtered, and few millimeters of the solution
were placed in the clean and dry watch glass just before the cell counting.
Before the main part of the exercise, clean and dry glass slides, sterile blood lancets, cotton,
70% alcohol, toothpicks, needles, anti-A and anti- B serum were prepared. This was done in
preparation for the drawing of blood which was used in the other part of the exercise.
A. Red Blood Cell Count
Blood was extracted in this part of the exercise. The finger tip of the subject was
wiped and clean using the 70% isopropyl alcohol and a cotton. The finger was then pricked with
a lancet, the first few drops of blood that comes out from the small wound was discarded or wiped
The rubber suction tube was attached to the upper end of the red blood cell pipette making
sure it was in a horizontal position while the mouthpiece was placed in the lips. The tip of the
pipette was placed in the drop of the blood making sure it stays at all times for the solid blood
column to fill the pipette. The rubber tube was sucked gently to draw blood until the 0.5 mark. The
procedure was done carefully to make sure that the pipette tip to never be detached from the blood
in order to avoid the formation of the air bubble inside the pipette. The tongue was placed against
the mouthpiece to hold the blood column and the excess blood was wiped out from the tip of the
pipette, the blood should be exactly at the 0.5 mark of the RBC pipette. The same pipette was used
to suck in the Hayem’s diluting fluid until the 101 mark. The rubber tubing was then removed and
both end of the pipette were covered by the thumb and the index finger to keep the liquid inside
the pipette intact while it was mixed in a circular motion for two minutes. Then the diluted blood
was poured into the clean and dry counting chamber, the first few drops of the blood solution was
discarded to avoid contamination. In pouring the diluted blood to the counting chamber, the cover
slip was first placed in the counting area, the tip of the pipette was placed in a position where its
tip is touching the edge of the cover slip on the hemocytometer. The fluid from the pipette was
released carefully by using the index finger to regulate the flow of the solution to the slip. After
two minutes of settling period the cell counting started with the counting using the microscope.
The counting starts at the cells in the 80 small squares on the five 1/25 mm2. The average
number of red blood cells per cubic millimeter of the whole blood was obtained and was identified
B. Agglutination
The finger of the subject was cleaned using the alcohol and cotton before it was punctured
with a sterile lancet. The first few drops of blood that came out from the wound was discarded
while the next following drops were collected in a clean and dry glass slide. The glass slide was
divided into the left and right portion. The left side labelled as A and the right side labelled as B.
Drops of blood was placed in both sides of the glass slide and anti-A serum was added in the blood
in the A side of the slide while anti-B serum was added in the B side of the slide. The blood and
For people with blood type A, coagulation should show after the anti-A serum was mixed
with the blood sample. Coagulation is also expected to form with the blood mixed with the anti-B
serum for people with a blood type B. coagulation should form when blood is both mixed with
anti-A and anti-B serum for people with a blood type-AB and no coagulation with both anti-A and
After the blood was mixed with the serum, coagulation was observed, and the blood type
A drop of blood was again collected from the same finger that was punctured for other
procedures. The blood was placed at the center of the clean and dry glass slide taking note of the
time that the blood was collected from the punctured finger. Using the tip of the lancet that was
used to puncture the finger of the subject was used to draw blood to the periphery to observe if any
fibrin clings to the tip of the lancet. The time was noted when fibrin formation was observed, and
Rats were put inside the container with a cotton with some drops of chloroform. Nine test
tubes were labeled A to I and was placed in the test tube rack accordingly. Blood were collected
from the already dead rats using a 2-cc syringe and a drop of blood was distributed to the nine
labelled test tubes. Test tube A contained only the blood sample and the clotting time was
determined. Test tube B was lined up with paraffin in which drop of blood was added. Test tube
C had cotton fibers at the bottom and sides of the test tube and again blood from the rat was also
placed in the test tube. Test tube D was placed in an ice bath right after the blood was dropped in
the tube while test tube E was placed in the water bath. 5 drops of 0.1% heparin was placed in test
tube F, small amount of 1% sodium oxalate in test tube G and small amount of 1% sodium citrate
in test tube H. The chemicals were dissolved in test tubes F, G and H when blood was added in the
tubes. Additional 1% calcium chloride was also mixed in the test tubes F, G and H, it was stirred
until the suspension of fine threads was observed. The test tube I with the blood sample was
continuously stirred, the clotting time of all the test tubes were identified except for test tubes F,
G and H.
Results and Discussion
Table 1. Blood type classification using the ABO blood typing system
Agglutination was observed when the blood sample was mixed with the anti-B serum. Blood type
O was also identified when there was no agglutination that happened when blood was mixed with
3.333333333, 3%
20, 20%
A
B
AB
63.33333333, 63% 13.33333333, 14%
O
The table above shows that most of the people in class has a blood O and only a few
The antigen-antibody interactions are the result of the agglutination which is the clustering
of the red blood cells in the liquid plasma. Antigens which are found in the red blood cell’s surface
is a biological signature of an individual’s blood type. The antigens are specific and are different
from other blood type. The antibodies in the blood recognize foreign blood types, which is why it
is necessary to test the blood’s compatibility before blood transfusion. When the antibodies
recognize the foreign antigens clumping of red blood cells occurs or the so-called agglutination
(O’Neil 1999).
Blood clotting occurs when the body tissue is damaged, it happens to prevent more
complicated damages, prevents severe physiological damages like blood loss. When the body
starts to bleed the coagulation cascade initiates the sequence of the clotting factor activities. The
clotting is mainly the function of the plasma and is mainly dependent on the interaction of the
plasma proteins, phospholipid, Ca++ and on the final stage it involves the interaction with the
thrombin which over all results to the conversion of soluble plasma protein fibrinogen to insoluble
fibrin which is what we observed in the laboratory. The blood contains receptors that binds with
the fibrin and later on attracts more platelets and fibrin to be able to form a clot. (Randall et all.
2002)
Figure 2. Theoretical Clotting Time in Different Conditions
The coagulation time is expected to be longer at the test tube lined with the paraffin wax,
compared to the empty test tube. One of the regulators of the conversion of prothrombin to
thrombin is released during the breakdown of the platelets to thromboplastin. The rate of the
coagulation depends on the said process and the coagulability and fluidity of blood is dependent
on the concentration of the said substances in the blood circulating the body (Donovan et al. 1949).
On the other hand, glass is identified as an activator of blood plasma which is also said to contribute
to the higher rate of coagulation in a glass surface. It is also said that thromboplastin increases in
a glass container this helps explain the longer time for the test tube lined with the paraffin (Lozner
et al. 1942). The activation of thromboplastin contributes to the higher coagulability of the blood
The blood clots also relatively faster in test tube with cotton fibers. The presence of the
fibers and some exposed glass surface agitates and activates the coagulation of blood (Tocantins
et al. 1951).
Stirring is said to make the clotting time slower or longer compared to set up where it was
just left to stand to clot. When stirring the fibrin adhere to the stirring rod thus preventing
Sodium citrate is one of the anticoagulants that prevents the coagulation of blood without
it altering the composition of the plasma. The sodium citrate removes calcium ions in the blood.
The calcium ion is essential in the coagulation process of the blood. The sodium citrate makes the
blood inactive thus no clotting will be observed. The sodium oxalate functions the same way with
the sodium citrate, it binds with the calcium in the blood thus forming insoluble calcium oxalate,
in this way calcium is not available thus clotting is not possible (Pal et al. 2005).
The blood clotting mechanism is much faster in higher temperature than in lower
temperature. Low temperature inhibits the effects on procoagulant enzymes which makes the
platelet disfunction which results to reduced platelet aggregation and this have a great impact on
the blood coagulation (Thorsen et al. 2011). On the other hand, blood agglutinates into clumps,
References:
fromhttp://anthro.palomar.edu/blood/blood_components.htm.
Randall, D., Burggren, W., & French, K. (2002). Eckert Animal Physiology, 5th edition. New
Lozner, E.L., Taylor, F.H.L., & MacDonald,H. (1942). The effect of foreign surfaces on blood
Tocantins, L.M., Carroll, R.T., & Holburn,R.H. (1951).The clot accelerating effect of dilution on
blood and plasma. Relation to the mechanism of Coagulation of normal and hemophilic
Pal G.K., & Pal, P. (2005).Textbook of Practical Physiology, Second Edition.Chennai, India:
Thorsen, K., Ringdal, K.G., Strand, K.,Soreide, E., Hagemo, J., & Soreide, K.(2011).Clinical and
98(7): 894-907.
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