Andrews University Biology Department Anatomy and Physiology II Spring 2008

Lab 1 - Blood and Blood Typing

Name _________________________ File # ________ TA Signature _______________

Introduction The study of the blood is termed hematology. Blood has three main functions: transportation, regulation, and protection. Blood supplies cells with oxygen and nutrients while removing carbon dioxide and other waste products. Specialized cells within blood protect us by fighting sources of infection and coagulation prevents uncontrolled blood loss when we get injured. The blood also safeguards body cells against other dangers such as extreme temperatures or acidity. Read Chapter 18 The Circulatory System: Blood, pages 679-712 (Saladin, 4th ed.) before coming to lab. IMPORTANT All items that have been in contact with blood must be placed in the biohazard bag right after you read your results.

Part I: ABO AND Rh DETERMINATION Red blood cells carry proteins on the erythrocyte (red blood cell or RBC) surface. In this lab exercise we will focus on proteins A, B, and D (Rhesus). These proteins can function as antigens (agglutinogens). Researchers have identified more than 100 erythrocyte antigens, each an expression of an inherited gene and each performing a specific function. An individual who is exposed to a foreign antigen (such as during an incompatible blood transfusion) may produce specialized proteins whose purpose is the destruction of the foreign antigen. These proteins, which are located in the serum fraction of the blood, are called antibodies. Once produced, an antibody can remain in the circulation for many years, ready to bind foreign antigens if introduced into the blood. Antibodies are specific; they will only bind to an antigen that is identical to the one that stimulated their production. In order to determine your ABO blood type, we will use antibodies present within anti-A and anti-B sera. If your blood type is A, the blood will agglutinate (clump) with the anti-A serum. It your blood is type B, it will agglutinate with anti-B serum. If your blood is type AB, it will agglutinate with both anti-A and anti-B sera. If your blood is type O (absence of proteins A and B), neither anti-A nor anti-B sera will cause agglutination. Human erythrocytes are also classified according to the Rh factor as either Rh positive (Rh +) or Rh negative (Rh -), depending on whether the Rh antigen is present or absent. This antigen is called the D-antigen. In order to determine the presence or absence of the Rh protein we will use anti-D

serum. Procedure: 1. Obtain a blood typing card for your ABO/Rhesus blood typing. Write your name on it. 2. Sterilize the tip of your right or left index finger with a sterile alcohol pad and use a sterile lancet to prick your finger. 3. Place one drop of blood onto each circular section of your blood-typing card. 4. Place one drop of anti-A serum on the first circular section of your card, one drop of anti-B serum on the middle section, and one drop of anti-D serum on the third section. 5. Thoroughly mix the blood and antisera with the end of a toothpick. Do not use the same toothpick for each sample! Use separate toothpicks for each section. After you are done mixing, place the toothpicks in the biohazard bag. 6. Observe for agglutination for the next 2 minutes. 7. Based on your observations, determine your ABO/Rhesus blood type and record your results below. (1 pt) Blood type ___________

Questions: 1. What antigens are present on a persons red blood cells if that person is: (6 pts) (a) type B+ (b) type O(c) type AB+

2. Explain the danger of giving a person with type B a blood a transfusion with type A blood. (5 pts)

3. In a paternity suit, a woman (type O) accuses a man (type A) of being the father of her baby (type O). Will the blood types prove or disprove her claim? Explain. (5 pts)

Part II: HEMOGLOBIN ESTIMATION USING THE TALLQUIST METHOD A mature erythrocyte is mainly a package of the molecule hemoglobin. This molecule gives the erythrocyte its oxygen-carrying power. If the concentration of hemoglobin in grams (g) per 100 milliliters (ml) is normal, the blood should have normal oxygen-carrying ability. Male values usually range from 13-18 g/100 ml, whereas female values normally are 12-16 g/100 ml. A departure from these norms, such as a deficiency, indicates that the oxygen-carrying capability is not normal. The amount of hemoglobin present in red blood cells is a good indicator of oxygen-carrying capacity of the blood. A simple method of measuring the amount of hemoglobin in blood is to compare a small piece of Tallquist paper that has been saturated with a sample of blood with a Tallquist color chart. Procedure: 1. Remove one square of paper from the Tallquist booklet. 2. Place 1-2 drops of blood on the center of the paper. Wait 15 seconds and compare your sample with the Tallquist scale. Hemoglobin concentration _________ g/dl (1 pt)

Part III: HEMATOCRIT Blood looks like a homogeneous red fluid. Closer analysis, however, reveals that the blood is composed of two basic parts: 1) the formed elements (cells) and 2) the plasma. The cells are the denser fraction, consisting of three major types: erythrocytes (red blood cells), leukocytes (white blood cells), and thrombocytes (platelets). The plasma generally constitutes a little more than half an adult humans 4-6 liters of blood. This fraction is 90% to 92% water, with various solutes dissolved or suspended within it. The percentage of erythrocytes found in a set volume of blood is known as the hematocrit or packed cell column (PVC). The adult male hematocrit normally averages 45%, with a range from 42% to 52%. The adult female hematocrit normally averages 42% and ranges from 37% to 48%. A low reading indicates possible anemia. Slightly higher levels may indicate a healthy adaptation. In this exercise you will calculate the hematocrit of a sample of blood.

Procedure: 1. Place a few drops of blood onto a slide. 2. Take a capillary tube and place one end of it on the blood at a 45-degree angle and let the tube fill up with blood about 3/4 of the way. 3. Seal the same end of the capillary tube that you placed in the blood with Seal-Ease. 4. Place the capillary tube into the microhematocrit centrifuge with the clay end facing the outside and put a tube filled with either with blood or water on the other side to balance. 5. After loading the tubes, tighten the inside and outside covers and set the timer for 2 minutes. 6. After 2 minutes take your tube out and place it in on the microhematocrit reader. Determine the percentage of blood volume from the scale. Hematocrit ___________ (1 pt)

Questions about hemoglobin and hematocrit tests: 1. Were your hemoglobin and hematocrit readings within normal limits? If not, give at least one reason why your hemoglobin and/or hematocrit levels deviate from the normal range (2 pts).

2. Give three reasons why normal hemoglobin and hematocrit values differ between males and females? (3 pts)