BLOOD TYPING

&
BLOOD SMEAR

LAB REPORT

NAME : NUR ADRIANA BT ZULKEFLEY

GROUP : 6A1
MATRIC NO : FAS 1704-7855
DATE : 1/3/2018
SUBJECT : INTRO TO ANATOMY & PHYSIOLGY (FG 1011)
INSTRUCTOR : MADAM SYAZAIRA ARHAM YAHYA ARIFF
OBJECTIVES :
 To state the characteristics and functions of the different types of blood cells.
 To explain the ABO and Rh blood group system.

INTRODUCTION :

 INTRO ON FORMED ELEMENTS OF BLOOD

- Human blood is made of different type of cells suspended in a solution called plasma.( 55% of
blood volume is plasma and 45% comprises of cells and solid constituents).
- Plasma is mostly water but contains many dissolved substances (gases, nutrients, wastes, ion,
hormones, enzyme, antibodies, and other proteins).
- Blood cells are formed in bone marrow of the long bones and lymph nodes.
- Erythrocytes (red blood cells or RBCs), Leukocytes (white blood cells or WBCs) and
Thrombocytes (platelets).
- Blood is the primary transport medium that is responsible for continuously supplying nutrients
and oxygen to the active cells in the body.
- Erythrocytes are tiny biconcave disks, and their primary function is to transport oxygen and, to a
lesser extent, carbon dioxide.
- Leukocytes provide a defense against organisms that cause disease and either promote or inhibit
inflammatory responses.
- Two main groups of leukocytes in the blood are granulocytes and agranulocytes; the cells that
develop granules in the cytoplasm are called granulocytes, and those that do not have granules are
called agranulocytes.
- Thrombocytes become sticky and clump together to form platelet plugs that close breaks and
tears in blood vessels.
- Three activities of the blood are transportation, regulation, and protection.
- The production of formed elements, or blood cells, is called hemopoiesis.
 INTRO ON BLOOD TYPING : A,B,AB AND O

- Antigens function to enable our immune system to distinguish the body’s own cells from foreign
invaders and destroyed by antibodies which produce in the immune system.
- Antigens on the surfaces of red blood cells (RBCs) are also known as agglutinogens and the
antibodies that react against them are also called agglutinins.
- People with type A blood have the antigen A on the surface of their RBCs.
- People with type B blood have the B antigen.
- Individuals with type AB blood have both A and B antigens.
- People with type O blood have no antigens present.
- The antibody that reacts against antigen A is called anti-A, and is found in the plasma of people
who do not possess antigen A on their own RBCs (people with type O or type B blood).
- The antibody that reacts against antigen B is called anti-B and is present in those who do not
possess antigen B in their RBCs (people with type O or type A blood).
- Agglutination is the clumping of RBCs bound together by antibodies. In giving transfusions, it is
very important that the donor’s RBCs not agglutinate as they enter the recipient’s bloodstream.

- Individuals either belong to the Rh-positive (Rh+) or Rh-negative (Rh-) Rhesus blood group.
- People with the Rh+ blood group have D antigens on the surface of their red blood cells, while
Rh- individuals do not.
- If an Rh- receives an Rh+ transfusion, the recipient produces anti-D antibodies.
- Since anti-D does not form instantaneously, there is little danger in the first mismatched
transfusion. However, because the recipient will now start producing anti-D, a subsequent Rh+
transfusion could agglutinate the donor’s RBCs.
MATERIALS ;

- Alcohol swab
- Sterilized lancet
- Anti-A , Anti- B, Anti- D serums
- An ABO/D board
- Wooden sticks
- Leishman stain

PROCEDURE :

 BLOOD TYPING

1. Wash your hands with soap and dry them off.Swing your hand for 10-15 seconds.
2. Apply alcohol swab to your middle finger and prick the tip of your middle finger using sterilized
lancet.
3. Wipe off the first blood drop.Place the next drop of blood in the middle of each box on the board
(ABO/D).
4. Place a drop of anti-A serum next to the box A blood cells,one drop of anti-B serum next to the
box B blood cells,and one drop of anti-D serum next to box D blood cells.Stir each set of boxes
with an unused mixing stick.
5. If the antigen present,the erythrocytes in that mixture will clump or the mixture will be cloudier.If
it isn’t present,the mixture will not change.
6. Record your blood type.

 BLOOD SMEAR

(a) Preparation of blood smear (Peripheral Blood Smear)

1. Place a small drop of blood, directly from freshly wiped fingertip puncture on 0.5 inch of
one end of clear side.
2. Using a second slide (spreader), place this slide slightly infront of the drop of blood.The
angle of this spreader slide must be approximately 45 ̊ .
3. Draw the spreader slide back toward the drop of blood.
4. Allow the drop of blood to spread about three-fourths of the way across the spreader
slide.This forward movement must be smooth and continue to the end of the slide.
5. A thin film of blood in the shape of bullet with a feathered edge will remain on the slide.
6. Allow the blood film to air-dry completely before staining.
(b) Blood smear staining

1. Cover smear completely with Leishman stain, flood the slide about 1-2 minutes.
2. Add equal volume of deionized water/phosphate buffer (pH 6.8).
3. Blow gently on slide to mix stain and buffer.Leave for 8-10 minutes.
4. Differentiate in buffe for 30 seconds.
5. Wash off and stand slides to dry.
6. Wipe off excess fluid from the back of the slide.Place the slide upright on a staining rack
with the feathered edge up and air-dry.
7. When completely dry,examine the smear with the microscope.
RESULTS
DISCUSSION :

1. Why do we need to swing our hands before pricking the tip of your finger?
- The inertia will allow the blood to flow down to the tip of the finger.

2. Why can’t you use the same lancet twice?

- Because the lancet is sterile to avoid microorganisms to reach your blood as bacteria and viruses.

3. Why do we need to wipe off the first blood tip?

- The reason is because the content of the blood gets in contact with the surface of the skin causing
any microorganism that are prevalent to be part of the sample.As well, the disinfectant used such
as alcohol will also get mixed with the sample taken and causes false interpretation of the results.

4. Why do you need different mixing stick to mix the blood and anti-serum on different boxes
on the slide?
- To prevent cross contamination.

5. Can an individual with O blood group donate his blood to a person with A blood
group?Explain
- Yes they can,because O blood group is a universal donor.

6. What information can we obtained from a patient’s blood film?

- The test provides information on the number and shape of these cells, which can help doctors
diagnose certain blood disorders or other medical conditions.

7. Explain how one can obtain a good peripheral blood smear?

- Three factors may be altered slightly to produce a perfect blood smear: speed, angle and drop
size.The faster the spreader slide is moved, the longer and thinner the film will be.The slower the
slide is moved, the shorter and thicker the slide will be.An angle greater than 30° makes the
smear thicker; less than 30° the smear is thinner.A small drop of blood may be insufficient to
prepare a slide of sufficient length, too large a drop may cause the smear to extend beyond the
length of the slide.

8. In a peripheral blood smear preparation,it is not advisable to have too large drop or too
small drop of blood on the slide,why?
- A small drop of blood may be insufficient to prepare a slide of sufficient length, too large a drop
may cause the smear to extend beyond the length of the slide.

9. List down the precautionary steps involved in this experiment.

- Make the blood smear without delay as soon as the drop of blood is placed on the glass slide, the
smear should be made.
- The spreader slide can only be used once to prevent carry over of patient’s blood on to the next
blood smear.
- Position your fingers on the spreader slide as far down as possible and apply even, moderate
pressure to the spreader slide. Any pressure exerted on the spreader slide should be directed
across the slide in the direction that the film is made rather than down on the stationary slide.

10. What are the characteristics of a well-made blood smear?

- Good smear is tongue shaped with a smooth tail.
- Does not cover the entire area of the slide.
- Has both thick and thin areas with gradual transition.
- Does not contain any lines or holes.

11. What are the features of a poor blood smear?

- Does not well spread.
- Contain air bubbles and holes.
- Cover the entire area of the slide.

12. What common mistakes can contribute to a poor blood smear?

- Drop of blood too large or too small.
- Spreader slide pushed across the slide in a jerky manner.
- Failure to keep the entire edge of the spreader slide against the slide while making the smear.
- Failure to keep the spreader slide at a 45° angle with the slide.
- Failure to push the spreader slide completely across the slide.

13. Describe and depict in a diagram the best optimal area in peripheral blood smear slide to
evaluate and enumerating blood cells.
- At the feathered edge where the blood smear do not the the edge of the slide.

14. What magnification is best use to evaluate the blood smear.Why?

- x 40 because the magnification is higher and the image seen is clearer and bigger.

15. Describe and depict in a diagram of the characteristic of red blood cell abnormalities.Give
three examples.
- Variations in size (anisocytosis)
- Variations in colour.
- Variations in shape of cell.
CONCLUSION
The purpose of this lab is to perform a slide ABO/Rh blood typing procedure, discuss the clinical
significance of ABO/Rh blood typing, and how antigens and antibodies relate to the blood typing
procedure. People can receive transfusions with only certain blood types, depending on the type of
blood they have. If incompatible blood types are mixed, erythrocyte destruction, agglutination and
other problems can occur. By using the ABO/Rh blood typing, one can find their blood type and
compatablity. Additionally, the ABO blood groups and other inherited antigen characteristics of red
blood cells are often used in medico-legal situations involving identification of disputed paternity.
Lastly,from the blood smear, we can identify and diagnose any abnormalities in our blood that causes
various types of diseases.

REFERENCES
- Introduction To Human Anatomy & Physiology Lab Manual.(FG 1011).(2018).
Experiment 3: Blood Typing & Blood Smear (page 1-9)
- EXERCISE 10: PREPARATION OF PERIPHERAL BLOOD SMEARS (2010).Chapter 10
Procedures for Collecting Capillary Blood Specimens.Retrieved from
http://www.austincc.edu/mlt/phb/phb unit9Lab10PreparationOfBloodSmearsFall2010.pdf
- Ofelia Gamez and Franky Lam.(2009). ABO/Rh Blood Typing Lab.Retrieved from
https://www.quia.com/files/quia/users/rjackso8/PPTs/Lab-8-Demonstation
- Lydia Krause.(June 6,2012).Blood Smear.Retrieved from
https://www.healthline.com/health/blood-smear