experiment 4 blood typing

Science in School

Investigating blood types teach article.

Author(s): Tim Harrison and Magdalena Wajrak

In this experiment, simple liquids that mimic blood are used to demonstrate blood typing.

The topic of blood types is often taught in school science lessons but experimenting with real blood may not be possible for many good reasons—because of the concerns of parents, the need for comprehensive risk assessments to prevent infection or the transmission of blood-borne disease, or the reluctance of students to use their own blood.

In this practical activity, simple chemical solutions are used to simulate blood types. The activity can be used in lessons, for a science club or as part of a forensic science day for students of many ages.

The science of blood

Blood is a sticky red fluid containing several kinds of cell suspended in a watery liquid called plasma: red blood cells, white blood cells and platelets (figure 1). Many chemicals are also suspended or dissolved in the plasma, including proteins, sugars, fats, salts, enzymes and gases. Each person’s blood has certain inherited characteristics that distinguish it from the blood of other people.

Until the 1980s, blood was primarily differentiated by ABO blood typing, which relies on the presence of three substances on the outside of red blood cells, called antigens. Although for forensic purposes, this technique has since been replaced by other methods such as DNA fingerprinting, for clinical purposes, ABO blood typing is still used before giving someone a blood transfusion to prevent complications such as rejection.

The presence or absence of A and B antigens on red blood cells determines a person’s ABO blood type. This leads to the identification of four main blood types: A, B, AB (when both antigens are present) and O (when neither antigen is present). A third important blood antigen is the Rhesus (Rh) factor, or D antigen. People with the D antigen are Rh positive, and those who lack it are Rh negative.

In order to type a person’s blood, antibodies (called agglutinins and sometimes referred to as antiserums) are added to a few drops of blood. These agglutinins bind to the antigens on the surface of the red blood cells, causing the cells to aggregate or clump. If clumping occurs in a blood sample, then that associated antigen is present. Once all antigens have been tested, the blood type can be deduced (table 1).

ABO blood typing experiment

Safety note.

Wear safety glasses and gloves. See also the general safety note .

Each group will need:

• Spotting tiles (dimple tray)
• Two pipettes, one for each blood sample
• 2.0 mol dm -3 hydrochloric acid solution in a dropping bottle labelled ‘Anti-A’
• 2.0 mol dm -3 sulfuric acid solution in a dropping bottle labelled ‘Anti-B’
• ‘O’ = distilled water
• ‘A’ = 0.1 mol dm -3 silver nitrate solution
• ‘B’ = 0.1 mol dm -3 barium nitrate solution
• ‘AB’ = a 50:50 mixture of 0.1 mol dm -3 silver nitrate and barium nitrate solutions
• Unidentified blood samples, made from the same solutions as the identified blood samples, labelled ‘victim 1’, ‘victim 2’, etc.

Explain the scenario to your students: there has been an accident and you need to know the ABO blood type of the victims before they can be given blood transfusions. It is the students’ job to use the blood samples and work out the type of blood each victim has.

• Using a clean pipette, put one drop of one of the identified blood samples into each of the first wells of the first two rows of your dimple tile. Complete the rows with the other blood samples, as shown in figure 2.
• Add a drop of Anti-A antiserum to each dimple in the first row and record your observations in table 2. If you’re not sure of a result, add another drop of Anti-A antiserum.
• Add a drop of Anti-B antiserum to each dimple in the second row and record your observations, also in table 2. If you’re not sure of a result, add another drop of Anti-B antiserum.
• Use your results to conclude how each blood type (O, A, B and AB) reacts to the antibodies.
• Take a clean dimple tray and test the victims’ blood using the same method. Record your observations in table 3.
• Use your results to assign the correct blood type to each victim.

What is happening?

These tests mimic how different blood types react with agglutinins, by using simple chemistry. With older students you may wish to discuss this chemistry, pointing out the differences between the antibody-antigen reaction which is being modelled and the simple displacement reaction that is actually happening.

In this experiment, instead of clumping blood cells, the (white) precipitates make the solutions clump in the spotting tiles.

Barium nitrate(aq) + sulfuric acid(aq) → barium sulfate(s) + nitric acid(aq)

Silver nitrate(aq) + hydrochloric acid(aq) → silver chloride(s) + nitric acid(aq)

The clumps that form are dark red, instead of white, because of the food colouring present.  

More types of artificial blood

The need to identify the blood type of patients before blood transfusion may soon be a thing of the past. Recently, UK-based researchers at the University of Edinburgh announced that they had made type O negative red blood cells from stem cells. If scaled up successfully, this method could lead to a new source of universal donor blood, and there are plans for a small-scale clinical trial in 2016.

Furthermore, researchers are developing products based on haemoglobin (the oxygen-carrying protein in blood), for example in a polymerised and powdered form that can be stored for months at room temperature, unlike blood, which has to be refrigerated.

Acknowledgement

There are a few alternatives to this experiment. The solutions used in this version came from a group of science technicians from the Association of Science Technicians in Independent Schools in Western Australia (LABNETWEST) w1 .

Web References

• w1 – To learn more about LABNETWEST visit their website .
• From the same website, why not explore why we have blood types at all?
• Wallace-Müller K (2011) The DNA detective game.  Science in School 19 : 30-35.
• Gardner G (2006) The detective mystery: an interdisciplinary foray into basic forensic science. Science in School 3 : 35-38.

Tim Harrison works at the University of Bristol, as the school teacher fellow at the School of Chemistry. This is a position for a secondary-school teacher that was created to bridge the gap between secondary schools and universities, and to use the resources of the School of Chemistry to promote chemistry regionally, nationally and internationally.

Dr Magdalena Wajrak is a chemistry lecturer and promotes chemistry with people of all ages. She is based at the School of Natural Sciences, Edith Cowan University, Perth, Australia.

This interesting practical activity addresses a basic topic of biology: blood types. Although the theory may be familiar to students, experiments on blood are normally avoided for reasons explained by the author. This, however, is an easy simulation to try in the laboratory using a simple chemical reaction.

The subject of this article could be related to other important topics such as immunology in biology, displacement reactions in chemistry, and even civil rights in ethics. The experiment could also be useful for awakening students’ interest in the need to investigate new materials and technologies that allow us to make safer and faster transfusions.

Suitable comprehension questions could include:

• What is the importance of knowing your blood type?
• Why does clumping occur? What is the relationship between antigen and antibody?
• Do you know an example of blood typing other than ABO?
• What is the difference between agglutination and displacement reactions?
• Why is it important to investigate new sources of universal donor blood?

Ana Molina, IES Gil y Carrasco, Ponferrada, Spain

Download this article as a PDF

Biology Lab Report- Blood Typing Lab

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Module 2: The Cardiovascular System: Blood

Blood typing, learning objectives.

By the end of this section, you will be able to:

• Describe the two basic physiological consequences of transfusion of incompatible blood
• Compare and contrast ABO and Rh blood groups
• Identify which blood groups may be safely transfused into patients with different ABO types
• Discuss the pathophysiology of hemolytic disease of the newborn

Blood transfusions in humans were risky procedures until the discovery of the major human blood groups by Karl Landsteiner, an Austrian biologist and physician, in 1900. Until that point, physicians did not understand that death sometimes followed blood transfusions, when the type of donor blood infused into the patient was incompatible with the patient’s own blood. Blood groups are determined by the presence or absence of specific marker molecules on the plasma membranes of erythrocytes. With their discovery, it became possible for the first time to match patient-donor blood types and prevent transfusion reactions and deaths.

Antigens, Antibodies, and Transfusion Reactions

Antigens are substances that the body does not recognize as belonging to the “self” and that therefore trigger a defensive response from the leukocytes of the immune system. (Seek more content for additional information on immunity.) Here, we will focus on the role of immunity in blood transfusion reactions. With RBCs in particular, you may see the antigens referred to as isoantigens or agglutinogens (surface antigens) and the antibodies referred to as isoantibodies or agglutinins. In this chapter, we will use the more common terms antigens and antibodies.

Antigens are generally large proteins, but may include other classes of organic molecules, including carbohydrates, lipids, and nucleic acids. Following an infusion of incompatible blood, erythrocytes with foreign antigens appear in the bloodstream and trigger an immune response. Proteins called antibodies (immunoglobulins), which are produced by certain B lymphocytes called plasma cells, attach to the antigens on the plasma membranes of the infused erythrocytes and cause them to adhere to one another.

• Because the arms of the Y-shaped antibodies attach randomly to more than one nonself erythrocyte surface, they form clumps of erythrocytes. This process is called agglutination .
• The clumps of erythrocytes block small blood vessels throughout the body, depriving tissues of oxygen and nutrients.
• As the erythrocyte clumps are degraded, in a process called hemolysis , their hemoglobin is released into the bloodstream. This hemoglobin travels to the kidneys, which are responsible for filtration of the blood. However, the load of hemoglobin released can easily overwhelm the kidney’s capacity to clear it, and the patient can quickly develop kidney failure.

More than 50 antigens have been identified on erythrocyte membranes, but the most significant in terms of their potential harm to patients are classified in two groups: the ABO blood group and the Rh blood group.

The ABO Blood Group

Although the ABO blood group name consists of three letters, ABO blood typing designates the presence or absence of just two antigens, A and B. Both are glycoproteins. People whose erythrocytes have A antigens on their erythrocyte membrane surfaces are designated blood type A, and those whose erythrocytes have B antigens are blood type B. People can also have both A and B antigens on their erythrocytes, in which case they are blood type AB. People with neither A nor B antigens are designated blood type O. ABO blood types are genetically determined.

Normally the body must be exposed to a foreign antigen before an antibody can be produced. This is not the case for the ABO blood group. Individuals with type A blood—without any prior exposure to incompatible blood—have preformed antibodies to the B antigen circulating in their blood plasma. These antibodies, referred to as anti-B antibodies, will cause agglutination and hemolysis if they ever encounter erythrocytes with B antigens. Similarly, an individual with type B blood has pre-formed anti-A antibodies. Individuals with type AB blood, which has both antigens, do not have preformed antibodies to either of these. People with type O blood lack antigens A and B on their erythrocytes, but both anti-A and anti-B antibodies circulate in their blood plasma.

Rh Blood Groups

The Rh blood group is classified according to the presence or absence of a second erythrocyte antigen identified as Rh. (It was first discovered in a type of primate known as a rhesus macaque, which is often used in research, because its blood is similar to that of humans.) Although dozens of Rh antigens have been identified, only one, designated D, is clinically important. Those who have the Rh D antigen present on their erythrocytes—about 85 percent of Americans—are described as Rh positive (Rh + ) and those who lack it are Rh negative (Rh − ). Note that the Rh group is distinct from the ABO group, so any individual, no matter their ABO blood type, may have or lack this Rh antigen. When identifying a patient’s blood type, the Rh group is designated by adding the word positive or negative to the ABO type. For example, A positive (A + ) means ABO group A blood with the Rh antigen present, and AB negative (AB − ) means ABO group AB blood without the Rh antigen.

The following chart summarizes the distribution of the ABO and Rh blood types within the United States.

In contrast to the ABO group antibodies, which are preformed, antibodies to the Rh antigen are produced only in Rh − individuals after exposure to the antigen. This process, called sensitization, occurs following a transfusion with Rh-incompatible blood or, more commonly, with the birth of an Rh + baby to an Rh − mother. Problems are rare in a first pregnancy, since the baby’s Rh + cells rarely cross the placenta (the organ of gas and nutrient exchange between the baby and the mother). However, during or immediately after birth, the Rh − mother can be exposed to the baby’s Rh + cells (Figure 1). Research has shown that this occurs in about 13−14 percent of such pregnancies. After exposure, the mother’s immune system begins to generate anti-Rh antibodies. If the mother should then conceive another Rh + baby, the Rh antibodies she has produced can cross the placenta into the fetal bloodstream and destroy the fetal RBCs. This condition, known as hemolytic disease of the newborn (HDN) or erythroblastosis fetalis, may cause anemia in mild cases, but the agglutination and hemolysis can be so severe that without treatment the fetus may die in the womb or shortly after birth.

Figure 1. The first exposure of an Rh − mother to Rh + erythrocytes during pregnancy induces sensitization. Anti-Rh antibodies begin to circulate in the mother’s bloodstream. A second exposure occurs with a subsequent pregnancy with an Rh + fetus in the uterus. Maternal anti-Rh antibodies may cross the placenta and enter the fetal bloodstream, causing agglutination and hemolysis of fetal erythrocytes.

A drug known as RhoGAM, short for Rh immune globulin, can temporarily prevent the development of Rh antibodies in the Rh − mother, thereby averting this potentially serious disease for the fetus. RhoGAM antibodies destroy any fetal Rh + erythrocytes that may cross the placental barrier. RhoGAM is normally administered to Rh − mothers during weeks 26−28 of pregnancy and within 72 hours following birth. It has proven remarkably effective in decreasing the incidence of HDN. Earlier we noted that the incidence of HDN in an Rh + subsequent pregnancy to an Rh − mother is about 13–14 percent without preventive treatment. Since the introduction of RhoGAM in 1968, the incidence has dropped to about 0.1 percent in the United States.

Determining ABO Blood Types

Clinicians are able to determine a patient’s blood type quickly and easily using commercially prepared antibodies. An unknown blood sample is allocated into separate wells. Into one well a small amount of anti-A antibody is added, and to another a small amount of anti-B antibody. If the antigen is present, the antibodies will cause visible agglutination of the cells (Figure 2). The blood should also be tested for Rh antibodies.

Figure 2. This sample of a commercially produced “bedside” card enables quick typing of both a recipient’s and donor’s blood before transfusion. The card contains three reaction sites or wells. One is coated with an anti-A antibody, one with an anti-B antibody, and one with an anti-D antibody (tests for the presence of Rh factor D). Mixing a drop of blood and saline into each well enables the blood to interact with a preparation of type-specific antibodies, also called anti-seras. Agglutination of RBCs in a given site indicates a positive identification of the blood antigens, in this case A and Rh antigens for blood type A + . For the purpose of transfusion, the donor’s and recipient’s blood types must match.

ABO Transfusion Protocols

To avoid transfusion reactions, it is best to transfuse only matching blood types; that is, a type B + recipient should ideally receive blood only from a type B + donor and so on. That said, in emergency situations, when acute hemorrhage threatens the patient’s life, there may not be time for cross matching to identify blood type. In these cases, blood from a universal donor —an individual with type O − blood—may be transfused. Recall that type O erythrocytes do not display A or B antigens. Thus, anti-A or anti-B antibodies that might be circulating in the patient’s blood plasma will not encounter any erythrocyte surface antigens on the donated blood and therefore will not be provoked into a response. One problem with this designation of universal donor is if the O − individual had prior exposure to Rh antigen, Rh antibodies may be present in the donated blood. Also, introducing type O blood into an individual with type A, B, or AB blood will nevertheless introduce antibodies against both A and B antigens, as these are always circulating in the type O blood plasma. This may cause problems for the recipient, but because the volume of blood transfused is much lower than the volume of the patient’s own blood, the adverse effects of the relatively few infused plasma antibodies are typically limited. Rh factor also plays a role. If Rh − individuals receiving blood have had prior exposure to Rh antigen, antibodies for this antigen may be present in the blood and trigger agglutination to some degree. Although it is always preferable to cross match a patient’s blood before transfusing, in a true life-threatening emergency situation, this is not always possible, and these procedures may be implemented.

A patient with blood type AB + is known as the universal recipient . This patient can theoretically receive any type of blood, because the patient’s own blood—having both A and B antigens on the erythrocyte surface—does not produce anti-A or anti-B antibodies. In addition, an Rh + patient can receive both Rh + and Rh − blood. However, keep in mind that the donor’s blood will contain circulating antibodies, again with possible negative implications. Figure 3 summarizes the blood types and compatibilities.

At the scene of multiple-vehicle accidents, military engagements, and natural or human-caused disasters, many victims may suffer simultaneously from acute hemorrhage, yet type O blood may not be immediately available. In these circumstances, medics may at least try to replace some of the volume of blood that has been lost. This is done by intravenous administration of a saline solution that provides fluids and electrolytes in proportions equivalent to those of normal blood plasma. Research is ongoing to develop a safe and effective artificial blood that would carry out the oxygen-carrying function of blood without the RBCs, enabling transfusions in the field without concern for incompatibility. These blood substitutes normally contain hemoglobin- as well as perfluorocarbon-based oxygen carriers.

Figure 3. This chart summarizes the characteristics of the blood types in the ABO blood group. See the text for more on the concept of a universal donor or recipient.

Chapter Review

Antigens are nonself molecules, usually large proteins, which provoke an immune response. In transfusion reactions, antibodies attach to antigens on the surfaces of erythrocytes and cause agglutination and hemolysis. ABO blood group antigens are designated A and B. People with type A blood have A antigens on their erythrocytes, whereas those with type B blood have B antigens. Those with AB blood have both A and B antigens, and those with type O blood have neither A nor B antigens. The blood plasma contains preformed antibodies against the antigens not present on a person’s erythrocytes.

A second group of blood antigens is the Rh group, the most important of which is Rh D. People with Rh − blood do not have this antigen on their erythrocytes, whereas those who are Rh + do. About 85 percent of Americans are Rh + . When a woman who is Rh − becomes pregnant with an Rh + fetus, her body may begin to produce anti-Rh antibodies. If she subsequently becomes pregnant with a second Rh + fetus and is not treated preventively with RhoGAM, the fetus will be at risk for an antigen-antibody reaction, including agglutination and hemolysis. This is known as hemolytic disease of the newborn.

Cross matching to determine blood type is necessary before transfusing blood, unless the patient is experiencing hemorrhage that is an immediate threat to life, in which case type O − blood may be transfused.

Critical Thinking Questions

• Following a motor vehicle accident, a patient is rushed to the emergency department with multiple traumatic injuries, causing severe bleeding. The patient’s condition is critical, and there is no time for determining his blood type. What type of blood is transfused, and why?
• In preparation for a scheduled surgery, a patient visits the hospital lab for a blood draw. The technician collects a blood sample and performs a test to determine its type. She places a sample of the patient’s blood in two wells. To the first well she adds anti-A antibody. To the second she adds anti-B antibody. Both samples visibly agglutinate. Has the technician made an error, or is this a normal response? If normal, what blood type does this indicate?
• In emergency situations, blood type O − will be infused until cross matching can be done. Blood type O − is called the universal donor blood because the erythrocytes have neither A nor B antigens on their surface, and the Rh factor is negative.
• The lab technician has not made an error. Blood type AB has both A and B surface antigens, and neither anti-A nor anti-B antibodies circulating in the plasma. When anti-A antibodies (added to the first well) contact A antigens on AB erythrocytes, they will cause agglutination. Similarly, when anti-B antibodies contact B antigens on AB erythrocytes, they will cause agglutination.

ABO blood group:  blood-type classification based on the presence or absence of A and B glycoproteins on the erythrocyte membrane surface

agglutination:  clustering of cells into masses linked by antibodies

cross matching:  blood test for identification of blood type using antibodies and small samples of blood

hemolysis:  destruction (lysis) of erythrocytes and the release of their hemoglobin into circulation

hemolytic disease of the newborn (HDN):  (also, erythroblastosis fetalis) disorder causing agglutination and hemolysis in an Rh + fetus or newborn of an Rh − mother

Rh blood group:  blood-type classification based on the presence or absence of the antigen Rh on the erythrocyte membrane surface

universal donor:  individual with type O − blood

universal recipient:  individual with type AB + blood

American Red Cross (US). Blood types [Internet]. c2013 [cited 2013 Apr 3]. Available from: http://www.redcrossblood.org/learn-about-blood/blood-types 2013.

• Anatomy & Physiology. Provided by : OpenStax CNX. Located at : http://cnx.org/contents/[email protected] . License : CC BY: Attribution . License Terms : Download for free at http://cnx.org/contents/[email protected]

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Science Projects > Life Science Projects > Blood Typing Science For Kids  

Blood Typing Science For Kids

While all blood contains the same components and looks pretty similar to the naked eye, human blood contains microscopic markers called antigens. And if incompatible blood is mixed, it can mean the difference between life and death. In this issue, learn about different blood types, as well as their accompanying compatibilities and incompatibilities.

Watch our video to see blood testing in action!

Human blood is made up of four main components: red blood cells, white blood cells, plasma, and platelets. Red blood cells, or erythrocytes, carry oxygen through the body and make up a little more than 44% of our blood content. White blood cells, or leukocytes, which are part of the immune system, fight disease by attacking and devouring harmful cells. Platelets, or thrombocytes, initiate blood clotting by pinching together damaged arteries. White blood cells and platelets together make up less than 1% of our blood. Liquid plasma, which makes up 55% of our blood, carries nutrients to the rest of the body.

Blood group types (A, B, AB, and O) were first discovered in 1900 by Austrian Karl Landsteiner, who was trying to understand why blood transfusion recipients often died. Blood typing is based on the way red blood cells clump together or agglutinate. Agglutination is caused by the presence of Anti-A and Anti-B antibodies reacting with the A and B antigens in the red blood cells. The red blood cells in Type A blood contain A antigens that clump in the presence of Anti-A antibodies but have no reaction in the presence of Anti-B antibodies. Alternatively, the red blood cells in Type B blood have B antigens that clump in the presence of Anti-B antibodies but do not react in the presence of Anti-A antibodies. Type AB blood contains both A and B antigens and will agglutinate with both Anti-A and Anti-B antibodies. Conversely, Type O blood contains neither A nor B antigens and will not react with either Anti-A or Anti-B antibodies. This is why people with type O blood are called universal donors, while people with blood type AB are universal recipients.

Blood Rh factor is another important blood grouping system that was discovered in the late 1930s. It gets its name because it was first found in the blood of rhesus monkeys. Blood can be either Rh negative or positive and simply indicates either the presence or absence of a specific antigen in the blood. Reaction with the Anti-D antibody shows a positive Rh factor. No reaction with the Anti-D antibody indicates a negative Rh factor.

If an expectant mother is Rh-negative and the father is Rh-positive, the unborn baby can inherit its father’s Rh factor. If the Rh-negative mother’s blood mixes with the Rh-positive baby’s, the mother’s body could develop antibodies against the baby’s blood. To protect the unborn baby, often the mother will receive an injection to prevent this. More than 85% of people are Rh-positive.</p><p>In summary, there are four main red blood cell types: A, B, AB, and O, and each of those can be positive or negative for the Rh factor. So in all, there are eight possible blood types: A+ or A-, B+ or B-, AB+ or AB-, and O+ or O-.

See our blood typing lab with 32 included blood tests and the Erycard Blood Type Test Kit .

Blood Compatibility Project *

Use this project to illustrate blood type compatibility. If the color of the “blood” changes, it is incompatible. If the color of the “blood” stays the same, it is compatible.

What You Need:

• 16 cups filled with water (four for each blood type)
• Red food coloring
• Blue food coloring
• Pen or pencil and paper to record data

What You Do:

1. Fill 16 cups with water.

2. Put red food coloring in four cups. They’ll represent Type A blood.

3. Put blue food coloring in four cups. These will represent Type B blood.

4. Put blue and red food coloring in four more cups to make a purplish color; this will represent Type AB blood.

5. Leave only water in the last four cups; this will represent blood Type O.

6. Pour one of the red “A” blood type cups into another one of the “A” blood type cups. Since the color did not change, blood Type A is compatible with blood transfusions with blood Type A. Once you’ve recorded that data, discard the cup.

7. Next, pour another red “A” into a blue type “B” cup. Since the color changed to purple, Type A blood and Type B blood are not compatible. Make a note of this as well.

8. Then pour a different “A” cup into the purple AB blood type.

9. Finally, red type A will pour the last cup into type O.

10. Repeat the steps with type B, AB, and O and record the results.

What Happened:

Blood Type A can only be given to Type A and AB patients. Blood Type B can only be delivered to Type B and AB patients. Blood Type AB individuals can receive blood from everyone, but can only donate to other AB blood type patients. Blood Type O individuals can only receive Type O blood, but they can donate blood to every other type.

*Project adapted from www.lessonplanspage.com

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