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Blood Laboratory Manual, Page 5

BLOOD LABORATORY MANUAL

Before coming to lab…

Read this report

Complete the Study Questions on the assignment worksheet

(these are the same questions in the teal-colored sections below).

Read the Laboratory Activity section – this will be completed during the laboratory session

Objectives:

How are red blood cell antigens and antibodies important in blood transfusions?

At the end of this Objective, you should also know:

A. How are antigens related to blood type?

B. How are antibodies used to determine blood type?

C. How do antigens and antibodies help us determine which blood types to use in transfusions?

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The figure on the left shows how you will mix 3 drops of patient’s blood with 3 different antibodies in the laboratory. In the part of the plate labeled A, you will add 3 drops of the blue anti-A antibody. In the part of the plate labeled B, you will add 3 drops of the yellow anti-B antibody. In the part of the plate labeled Rh, you will add 3 drops of the green anti-Rh antibody. The figure on the right shows what we would expect to happen if the patient had blood type A+. There would be an agglutination reaction with the anti-A antibody when it binds to the A antigen on the surface of these red blood cells. There would also be an agglutination reaction with the anti-Rh antibody when it binds to the Rh antigen on the surface of these red blood cells.

Study Tip: While you read these notes, make your own drawings or models of red blood cells, antigens, and antibodies.

Clinical Application:

There is a disaster, and many people need blood transfusions. For some people, their same blood type is not available. Which other blood types can they receive?

What Do You Already Know About…

(These questions are designed to get you thinking about what you already know so you can connect your learning below to previous knowledge as this is how we learn.)

· the parts of blood and the functions of each of these parts?

· red blood cells?

· blood typing and blood transfusions?

· antigens and antibodies?

· Rh antigens during pregnancy?

What are the parts of blood?

Blood has two main parts: (1) plasma and (2) cellular components. (1) Plasma is mostly water. Plasma also contains gases such as oxygen and carbon dioxide, nutrients such as glucose and amino acids, and proteins such as antibodies and clotting factors.

(2) Cellular components include three main types: (a) platelets, (b) white blood cells, and (c) red blood cells. (a) Platelets are cell fragments from a larger cell and function in clotting. (b) White blood cells come in many types, and they function in inflammation and immunity to help us to respond to injury including pathogens.

(c) Red blood cells (RBCs) are the most abundant cellular component in blood and make up about one half of your blood. RBCs are flexible cells which are roughly the size of capillaries. During development, RBCs lose their nuclei, and they do not have mitochondria.

The main function of the red blood cells is gas exchange. RBCs help transport oxygen from the lungs to body cells and carbon dioxide waste from your cells to your lungs for exhalation. Hemoglobin within RBCs binds to these gases to help with transport.

Study Questions:

(These questions are designed to get you thinking about the provided material. These are the same questions as on Part 1 of the blood assignment worksheet that you will complete before the laboratory session by the due date. Type your answers to these questions on that worksheet).

(1) What at the main parts of blood and a function for each?

How are antigens and antibodies involved in blood transfusions?

If you have a problem with your red blood cells, you might need a blood transfusion. But, before you can get someone else’s blood cells, you need to make sure your donor blood is a match. What does this mean?

Remember that molecules in our bodies have specific shapes, and that these shapes can be recognized by the shapes of other molecules. For example, this is how neurotransmitters and drugs bind to receptors and how enzyme active sites bind to substrates.

These molecular shapes are also recognized by molecules of our immune system, such as antibodies. Our immune system helps us determine what should be in our body and fight off things that are foreign in our bodies.

An antigen is any molecule that can be recognized by the immune system.

· We often think of antigens on microorganisms such as bacteria or viruses. These are foreign antigens.

· Our bodies are full of three-dimensional molecules. These are self-antigens.

One example of antigens are the glycoproteins on red blood cells for example, the A and B antigens). The antigens on your red blood cells are your self antigens. If you receive a blood transfusion, the antigens on the donor blood are foreign antigens. You want these to “match” so that your immune system does not destroy the donated blood.

Antigens on the outside of cells can identify cells to other cells and molecules. You might think of an antigen as an address outside of a house.

Antibodies are proteins made by a type of white blood cell called a B cell (also called a B lymphocyte). Antibodies are secreted from B cells and circulate in the blood. When foreign antigens enter the body (for example, on a virus), antibodies can bind to the foreign antigens to help the immune system destroy them. So, antibodies and other molecules in the immune system help the body get rid of foreign antigens. This helps us fight infections.

However, if our antibodies and other immune system molecules bind to our self antigens, the immune system destroys our normal cells. This is what happens in autoimmune diseases (auto means self).

We have a way to keep our immune system from destroying our body cells. If immune cells react with our self antigens, these cells are killed. For example, if a B cell antibody reacts with an antigen on the outside of your red blood cells, this B cell would be killed, and then these self-reacting antibodies would be removed.

Cells that do not react with our self-antigens survive and can then react with foreign antigens. This allows our immune cells to tell the difference between antigens on our own cells from foreign antigens that need to be removed from the body. This is called immune tolerance, or the ability to distinguish self from non self.

Each person has unique antigens. If a beneficial foreign tissue, such as an organ, tissue, graft, blood, or fetus, is given to another person, the foreign antigen one the donor cells will be recognized as foreign by the recipient’s immune cells unless it is an exact match (the only exact match is an identical twin).

Study Questions:

(2) In your own words, what is an antigen?

(3) In your own words, what is an antibody?

(4) What is one way that antigens and antibodies are similar?

(5) What is one way that antigens and antibodies are different?

Why are our blood types important?

There are antigens on the outside of our red blood cells. The ones hear about most often are the A, B, O, and Rh antigens. These are what are matched during blood transfusions. Since they are inherited in a predictable way much like sickle-cell anemia, they can also be used to help determine paternity (see below).

The A, B, O, and Rh antigens are glycoproteins on the surface of red blood cells. The O antigen is one type of glycoprotein in nearly every person. The A antigen is more carbohydrate added to this O antigen. The B antigen is a different carbohydrate added to this O antigen. Many people think of O as the absence of antigen, and since this is the scaffold for other antigens and in nearly all people, this is an okay assumption.

A person who…

· has the A antigen has A blood

· has the B antigen has B blood

· has the A and B antigen has AB blood

· has neither the A or the B antigen has O blood

· also has the Rh antigen has positive (+) blood so would be A+, B+, AB+, or O+

· does not express the Rh antigen has negative (-) blood and would be A-, B-, AB-, or O-

Antigen DescriptionAntigen NameAnalogyBlood Type
Antigen on red blood cells in nearly every person“O” antigenA room number outside of every office door in a buildingO
Sugars linked to the O antigenA antigenPictures (and room numbers) outside of office doorsA
Different sugars linked to O antigenB antigenAnnouncements (and room numbers) outside of office doorsB
A and B sugars linked to O antigenA and B antigensPictures and announcements (and room numbers) outside of office doorsAB
Another molecule on red blood cellsRh antigenA light (and room numbers) outside of office doors (O+). There could also be pictures (A+) or announcements (B+) or pictures and announcements (AB+)Rh+(or Rh-)

For example, if you have A+ blood, this means your red blood cells have A antigens and Rh antigens on the surface. When your immune cells develop, B cells making antibodies that react with A or Rh antigens will be destroyed. So, you will not make antibodies that react with A or Rh antigens (anti-A antibodies or anti-Rh antibodies). You could receive the following blood types: A-, A+, O-, O+.

But, since your red blood cells do not have B antigens, it is okay if you made antibodies that bind to B antigens. If you then received red blood cells with B antigens on the surface, your anti-B antibodies would see these cells as foreign and kill them. This is called hemolysis (hemo = blood, lysis = bursting). In other words, you could not receive B+, B-, AB+ or AB- blood.

Study Questions:

(6) If you have B- blood, which antigens are on the surface of your red blood cells?

(7) If you have B- blood, which antibodies should you NOT make due to immune tolerance?

(8) If you have B- blood, which antibodies could your body make?

(9) If you have B- blood, which blood types could you receive in a transfusion?

(10) If you have B- blood, which blood types would you NOT want to receive in a transfusion?

How are blood types inherited?

Remember that genes are cellular instructions, so each of the antigens on RBCs has a gene. Also remember that we usually have two copies of each gene, one from our mother and one from our father. We call these two copies alleles.

Often genes/alleles are dominant or recessive. Dominant genes/alleles “dominate” and one are expressed. If a person inherits one dominant and one recessive gene, then the dominant gene dominates and is the one that is seen in the cell or person. To have the traits of recessive genes/alleles, you need to have two copies of the recessive gene/allele.

The Rh antigen is expressed in this way. If a person has at least one dominant allele for Rh factor, he/she will express the Rh antigen on their red blood cells, and he/she will be Rh positive. If a person has two recessive alleles for Rh factor, he/she will not express the Rh antigen on red blood cells, and he/she will be Rh negative.

Study Questions:

(11) If a woman is Rh negative, will she be able to make anti-Rh antibodies? Why or why not?

The genes for blood type are not dominant or recessive. The ABO antigens are co-dominant. Both genes/alleles are expressed. Basically, there are three alleles: A, B, O.

A person who…

· has A blood is either AA or AO

· has B blood is either BB or BO

· has AB blood is AB

· has O blood is OO

For example, if a woman with AB blood mates with a man with O blood, there is a 50% chance the children will have A blood and a 50% chance the children will have B blood.

AB
OAOBO
OAOBO

Study Questions:

(12) Can a woman with A blood mate with a man with B blood and have children with O blood?

(13) Which genes would the man and woman have in the example in the question above?

Why is Rh important during pregnancy?

Rh antibodies can cross the placenta. Therefore, it can become a problem if a woman is Rh negative is pregnant with an Rh-positive fetus.

If the mother is Rh negative, she can make anti-Rh antibodies that react with Rh antigens. The mother’s anti-Rh antibodies could cross the placenta and react with the Rh antigens on the fetal red blood cells and destroy them. This is called erthyroblastosis fetalis (erthyro = red, blast = immature cell, osis = condition of). Since the mature red blood cells are destroyed, immature red blood cell blasts are released from bone marrow.

If an Rh negative person is exposed to Rh antigens, it takes awhile to produce the anti-Rh antibodies. Therefore, during a first transfusion or pregnancy, there is usually not a problem. However, during another transfusion or pregnancy, there can be a reaction.

If an Rh negative woman is pregnant with an Rh positive fetus, she is given RhoGam which is anti-Rh antibodies. The RhoGam destroys Rh-positive fetal blood in the mother’s circulation so she does not produce her own anti-Rh antibodies.

Study Questions:

(14) There was a crisis situation. You have two patients, one male and one female, who each have A- blood. Both patients are in critical condition, and each one needs one pint of blood. You have one pint of A- blood and one pint of O+ blood. Which would you give to which patient and why? (Hint: which patient is more likely to have an exposure to Rh antigen before or after the transfusion?)

How will we determine blood types in the laboratory?

Laboratory Activity:

(This will be completed during the laboratory session – see picture at the start of the Objective. Read it and be ready to perform these steps.)

1. Each person in your team should obtain one blood-typing plate and the blood of one “patient.”

2. Each team should have at least one set of antibodies: a bottle of blue anti-A, a bottle of yellow anti-B, and a bottle of green anti-Rh.

3. Each person in your team should add 3 drops of the patient blood to each of the wells in the blood-typing plate.

4. Each person should then add 3 drops of the blue anti-A antibodies to the testing-plate well labeled A, 3 drops of the yellow anti-B antibodies to the testing-plate well labeled B, and 3 drops of the anti-Rh antibodies to the testing-plate well labeled Rh.

5. Shake or swirl the plate to mix. It may take a few minutes to see the reaction. A positive reaction will look cloudy and/or like there is lint or dust in the well. You are observing agglutination, or clumping, of the red blood cells when the antibodies links the red blood cells together. Record the reactions as directed in laboratory.

6. As soon as you are finished observing the reaction, rinse out the plates at the sinks at the side of the room and leave the clean plates on the paper near the sinks. Do not let the reactions to sit on your bench for extended periods of them as this makes and ruins the plates.

Where can I learn more? (This last page is not required and is provide if you need more help.)

Watch: Animation This optional video with text/captions will help you relate antigens on red blood cells to antibodies and reactions (this video is showing Rh-negative cells).

Available at http://highered.mcgraw-hill.com/sites/0072495855/student_view0/chapter21/animation__cytotoxic__type_ii_hypersensitivity_.html

Read: Blood Types. This optional web page with charts will help you relate blood types to antigens, antibodies, blood donation, and inheritance.

Available at http://www.redcrossblood.org/learn-about-blood/blood-types

Read: What is a blood type? This optional web page with pictures will help you compare and contrast blood types, antigens, and antibodies (look for patterns).

Available at http://www.nobelprize.org/educational/medicine/bloodtypinggame/1.html

Read: How do you determine a person’s blood type? This optional web page with pictures will help you to relate laboratory blood testing to blood types.

Available at http://www.nobelprize.org/educational/medicine/bloodtypinggame/2.html

Read: How do you perform safe blood transfusions? This optional web page with pictures and charts will help you match patient blood types to donor blood types during transfusions.

Available at http://www.nobelprize.org/educational/medicine/bloodtypinggame/3.html

Play: The Blood Typing Game. This optional game from the Nobel Prize web site will help you predict which blood types patients can receive during transfusions (choose any of the 3 versions).

Available at http://www.nobelprize.org/educational/medicine/bloodtypinggame/game/

Play: Blood Typing Game. This optional game from the Red Cross web site will help you match patient blood types to donor blood types during transfusions.

Available at http://www.redcrossblood.org/donating-blood/donor-zone/games/blood-type

Read: Blood Typing Overview. This optional web page with pictures will help you to relate antigens, antibodies, and laboratory blood testing to blood types.

Available at: http://waynesword.palomar.edu/aniblood.htm

Read: Famous Black Inventors – Dr. Charles Drew Blood Bank Inventor. This optional reading with links will help you understand how this researcher helped to understand plasma so that blood could be stored.

Available at: com/Dr-Charles-Drew.asp”>http://www.black-inventor.com/Dr-Charles-Drew.asp

The Big Picture:

· The main function of RBCs is to transport oxygen to cells and help remove carbon dioxide.

· Antigens bind to molecules and cells in our immune system.

· Antibodies are proteins made by cells in the immune system that can bind to antigens.

· Immune cells that bind with our self-antigens are killed.

· Molecules on the outside of our red blood cells determine our blood type.

· Antibodies can react with antigens on red blood cells and cause the immune system to kill them.

· During blood transfusions or pregnancy, red blood cells are killed if we make antibodies that react with antigens on donor or fetal blood.

Integration and Application:

(These questions are provided to help you review this material. You do not need to turn these in.)

(1) What is the main function of red blood cells and hemoglobin?

(2) How are antigens similar and different?

(3) Why don’t we react with our own antigens and red blood cells?

(4) Which antigens are on the outside of red blood cells?

(5) Which antibodies react with which blood types?

(6) For each blood type, which antibodies can be circulating in the blood? Which cannot?

(7) How are antigens on the outside of red blood cells important in blood transfusions?

(8) For a person with each blood type, which blood types can be donated?

(9) How are blood types inherited?

(10) Why is Rh important in pregnancy?

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Blood Assignment

Part 1: Blood Introduction Study Questions

Read the Blood Introduction posted online and answer the Study Questions below (these are the same questions as within the provided introduction document).

(1) What at the main parts of blood and a function for each?

(2) In your own words, what is an antigen?

(3) In your own words, what is an antibody?

(4) What is one way that antigens and antibodies are similar?

(5) What is one way that antigens and antibodies are different?

(6) If you have B- blood, which antigens are on the surface of your red blood cells?

(7) If you have B- blood, which antibodies should you NOT make due to immune tolerance?

(8) If you have B- blood, which antibodies could your body make?

(9) If you have B- blood, which blood types could you receive in a transfusion?

(10) If you have B- blood, which blood types would you NOT want to receive in a transfusion?

(11) If a woman is Rh negative, will she be able to make anti-Rh antibodies? Why or why not?

(12) Can a woman with A blood mate with a man with B blood and have children with O blood?

(13) Which genes would the man and woman have in the example in the question above?

(14) There was a crisis situation. You have two patients, one male and one female, who each have A- blood. Both patients are in critical condition, and each one needs one pint of blood. You have one pint of A- blood and one pint of O+ blood. Which would you give to which patient and why? (Hint: which patient is more likely to have an exposure to Rh antigen before or after the transfusion?)

Part 2: Blood Laboratory Table

Complete the last four columns by:

1. deleting incorrect options or

2. highlighting correct options.

BloodTypeAntigensIf you mixed these RBCs with antibodies, which antibodies would react with antigens on the surface of the RBCs?(Which antibodies would react in a blood typing reaction?)A person with this blood type could have which antibodies in their blood?(Which antibodies would not react with the antigens on the surface of the RBCs?)If a person with this blood type needed a blood transfusion, which blood type(s) could they receive?
O-ABRhanti-Aanti-Banti-Rhanti-Aanti-Banti-RhO- O+A- A+ B- B+AB- AB+
O+ABRhanti-Aanti-Banti-Rhanti-Aanti-Banti-RhO- O+A- A+ B- B+AB- AB+
AB-ABRhanti-Aanti-Banti-Rhanti-Aanti-Banti-RhO- O+A- A+ B- B+AB- AB+
AB+ABRhanti-Aanti-Banti-Rhanti-Aanti-Banti-RhO- O+A- A+ B- B+AB- AB+

Part 3: Blood Note Card

1. Choose one term: antigen or antibody

2. Prepare one note card for that term using the criteria below.

For this assignment, “note card” is defined loosely and could be a 3×5 or 4×6 card or a whole sheet of paper. To submit it electronically, you can type up your note card using the next page as a worksheet. Or, you could make your own cards then take a picture or scan a note card.

You will earn 0.2 points for each of the 10 items listed for the “back” of each note card. Please number each part. See posted example.

The Big Ten (things to put on study cards):

Front:

the term itself, or the principle or the concept, spelled correctly

pronunciation, if necessary

where did you find this information? (textbook page, web link, citation)

Back:

1. the definition in your own words

2. an example from the class materials or another source

3. how it works

4. if something causes it to increase, what effect will it have?

5. if something causes it to increase, what effect will it have?

6. what it is a part of (in which larger structure or category can it be found)

7. what is part of it (which smaller structures or concepts are found in it)

8. what it is similar to

9. what it is different from (what it isn’t)

10. drawing, chart, mnemonic device, analogy, connection to something in your life, etc.

In Physiology, you will be expected to answer questions testing higher-order thinking skills such as predicting and comparing. To do this on a test, you need to practice while studying. This note card is one way to help you do this.

· It is not expected that you will use all of these items on your own cards, but think of the ones that were helpful to you, and include them on future cards.

· Take a stack of note cards with you (or study guides you have prepared) to use when you have short breaks in your schedule (for example, waiting for work or class to start or when picking up your kids, making dinner, on the bus, etc.)

· Use your note cards to see connections between concepts. For example, you could use cards to construct concept maps or pathways. Once you make a pathway, ask yourself what will happen if one thing in the pathway increases or decreases.

· Use your note cards to quiz yourself beyond definitions. For example, ask yourself

· what happens if it increases or decreases

· what comes before it or after it in a pathway

· what happens to it if the step before it increases

· what it is similar to, what it is different from

Part 3: Blood Note Card Worksheet

Front:

Term: ________________

Pronunciation:

Where did you find this information? (textbook page, web link, citation)

Back:

1) the definition in your own words

2) an example from the class materials or another source

3) how it works

4) if something causes it to increase, what effect will it have?

5) if something causes it to decrease, what effect will it have?

6) what it is a part of (in which larger structure or category can it be found)

7) what is part of it (which smaller structures or concepts are found in it)

8) what it is similar to

9) what it is different from (what it isn’t)

10) drawing, chart, mnemonic device, analogy, connection to something in your life, or other method you will use to help you remember it