Lab 6 Blood Exercise 29 Blood Analysis: Computer Simulation Exercise 29B PhysioEx or PhysioEx Chapter 17 in Mastering A&P Blood Activity 2: Examining the Formed Elements of Blood Microscopically, 6 (Using a prepared slide.) Other Directions in Handout Blood Analysis - PhysioEx Activity 1: Hematocrit Determination, 1 - 10 Activity 5: Total Cholesterol Determination, 1 – 10 Use directions in the lab manual or the handout at the front desk. Lab 6 Name___________________________ Lab Section _____________ Draw a picture of each type of blood cell, using the characteristics you described in the pre-lab exercise. Color each cell appropriately. Be sure that you use the correct relative sizes for each cell. Label nucleus and granules if appropriate. Colored pencils are available in the lab. (26 points) Activity 3: Prepared Slide of Blood Smear Use a prepared slide of a blood smear and be sure you can identify all the formed elements on a slide. Use the photos in the lab manual histology atlas or on web sites, and your dichotomous key to help you identify the cells. Use the 40X lens. Activity 3(Modified): Conducting a Differential WBC Count. Differential White Blood Cell Counts (Also Called CBC – Complete Blood Count) (Modified from http://www.rnceus.com/) Normal values for total WBC and differential in adult males and females are: Total WBC: 4,500 - 10,000/mm3 Bands (immature neutrophils): 3 - 5 % Granulocytes (or polymorphonuclears) o Neutrophils: 50 - 70% relative value (2500-7000 absolute value) o Eosinophils: 1 - 3% relative value (100-300 absolute value) o Basophils: 0.4% - 1% relative value (40-100 absolute value) Agranulocytes o Lymphocytes: 25 - 35% relative value (1700-3500 absolute value) o Moncytes: 4 - 6% relative value (200-600 absolute value) Each differential always adds up to 100%. To make an accurate assessment, consider both relative and absolute values. For example a relative value of 70% neutrophils may seem within normal limits; however, if the total WBC is 20,000, the absolute value (70% x 20,000) would be an abnormally high count of 14,000. The absolute value is the % of a cell type times the total WBC count. Leukocytes: critical low and high values Leukocytosis (high WBC count): a WBC count above 10,000 A WBC count over 30,000 indicates massive infection or a serious disease such as leukemia. Leukopenia(low WBC count): WBC count falls below 4,000 A WBC count of less than 500 places the patient at risk for a fatal infection. 1. You will be given a card with 50 WBCs on it. It will also contain information about the total WBC count for the sample and other information if relevant. Count the number of each of the different types of WBCs and fill in the chart below. Multiply your counts by 2 to find the differential WBC count for your sample. (The differential count is based on 100 cells and is reported as a %) 2. Using your data, decide which of the following eight scenarios is the most likely explanation for your results. Scenarios This patient has AIDS. His lymphocyte count is low, which makes the percentages of other WBCs seem high because the total WBC count is in the normal range. This patient has acute appendicitis, caused by a bacterial infection. Her total WBC count is high and her neutrophil count is high as well. There is a higher than normal percentage of “bands” in the sample. This patient has viral mononucleosis. Her lymphocyte count is high as lymphocytes enter her blood to fight the viral particles. The lymphocytes look a bit atypical with ragged edges. This patient just returned from the tropics and has picked up a filarial (roundworm) infection. His eosinophil count is high as the eosinophils attempt to fight off the parasite infection. This patient has acute myeloid leukemia. Red blood cell and platelet levels are low as are the WBC levels. Patient E is tired, achy and bruises easily. This patient has swollen lymph nodes and a high lymphocyte count. He is suffering from B-cell chronic lymphocytic leukemia (B-cell CLL). This patient is feels tired, is gaining weight, is very sensitive to the cold and has an elevated basophil count. She has been diagnosed with hypothyroidism. This patient has tuberculosis which, among other things, has resulted in a condition called monocytic leukocytosis or monocytosis. Cells Segmented neutrophils Band neutrophils Tally Total Lymphocytes Monocytes Eosinophils Basophils Total cells counted ___________ What is your patient’s total WBC count? (This is on the card.) ____________________ Is this normal, high or low? ____________________ What is the % of each of the cell types in the sample? This is the tally x2 if there are 100 cells. In some cases there are a few more that 100 cells. Just multiply the count by 2 as it will not make a major difference. This number represents the % of each blood cell type. Record in the table below. What is the absolute value of each of the cell types you counted? This is the % of each cell type x the total number of cells. Record in the table below. Normal Counts Segmented Neutrophils 50-70% Band Neutrophils 05% Lymphocytes 2540% Monocytes 3-8% Eosinohils 2-4% Basophils 0-1% Your Patient’s WBC Type Count (%) (Tally X 2) Your Patient’s Absolute Count Evaluation High, Low, Normal Compare your counts with the normal percentages and diagnose your patient’s condition. Your counts are percentages because they are based on 100 WBCs. Put the letter of your patient here ________ What is your diagnosis and why? ________________________________________________________________________ ________________________________________________________________________ _______________________________________________________________________ Activity 7: Blood Typing ( modified) (From Ward's or Carolina’s Simulated ABO and Rh Blood Typing) Blood typing is an example of an antigen-antibody reaction. Antibodies are a normal part of the immune response. In this example, the antigen-antibody complex that forms is a clump of blood cells. A. Refer to your text, page 671, or the lab manual, pages 433-435 for information about ABO and Rh blood groups and blood typing. B. Obtain a blood typing kit for your lab group. The kit should contain: 1 vial of simulated Anti-A Typing Serum 1 vial of simulated Anti-B Typing Serum 1 vial of simulated Anti-Rh Typing Serum 4 vial of blood samples 1 blood typing tray toothpicks glass marking pencil C. Using the wax marking pencil, label each blood typing slide with the number of the sample to be tested (1, 2, 3, 4). D. Follow directions for the Ward’s Carolina kit. (Circle one) Carolina: Place one drop of simulated blood in each well of the typing slide. Add one drop of anti-A serum in each A well Add one drop of anti-B serum in each B well Add one drop of anti-Rh serum in each Rh well Stir each well with a different clean toothpick. Avoid cross contamination. Wait a few minutes and observe each well. Record observations on the data sheet, and answer the related questions. Ward’s: Place three or four drops of simulated blood in each well of the typing slide. Add three or four drops of anti-A serum in each A well Add three or four drops of anti-B serum in each B well Add three or four drops of anti-Rh serum in each Rh well Stir each well with a different clean toothpick. Avoid cross contamination. Wait a few minutes and observe each well. Record observations on the data sheet, and answer the related questions. Blood Typing Results A. Record your clumping (agglutination) observations below: use + to indicate clumping use - to indicate no clumping Sample # Anti-A Anti-B Anti-Rh Serum Serum Serum Blood Type 1 2 3 4 B. Questions (Check your results with me before proceeding with the questions.) Answer these questions at home after you have completed the lab. They will be useful in preparing the poster for a poster session next week. 1. Give a general definition of antigen. 2. Give a general definition of antibody. 3. What ABO and Rh antigens are present on the surface of each sample of blood? Sample # Blood Type ABO antigens A, B Rh antigens ( + or - ) 1 2 3 4 4. For several possible reasons, blood serum contains antibodies to ABO antigens NOT present on the red blood cells in that blood. (The same is not immediately true for Rh antigens.) What ABO antibodies are in the serum of these blood samples? Sample # Blood Type ABO antibodies in the serum A, B 1 2 3 4 5. If any of these people needed a transfusion, what ABO blood type(s) could be used? Sample # 1 2 3 4 Blood Type ABO antigens on RBCs ABO antibodies in the serum ABO Blood Types for transfusions What happens to erythrocytes if incompatible ABO blood types are mixed? Which of the people should not be given Rh+ blood? _________________ Why not? 6. Answer the following questions based on the results from Sample 3. What is the ABO and Rh genotype of this person? _________ phenotype? ___________ Can a man with Type O blood be the father of this person? _______ Why or why not? (Illustrate your answer using a Punnett Square – ABO blood groups only.) Can this person have one parent with Type A blood and one with Type B blood? _____ Why or why not? (Illustrate your answer with one example on the Punnett Square – ABO blood groups only.) Can this person be the parent of a Type O child? _______ Why or why not? Illustrate your answer using the Punnett Square.) This person has parents who are both Rh+. How is this possible? (Illustrate your answer on a Punnett square.) Exercise 29B PhysioEx, page PEx 65 or on Mastering A&P Chapter 17 Blood Analysis: Total Cholesterol Determination Activity 1: Hematocrit Determination, 1 - 10 Activity 5: Total Cholesterol Determination, 1 - 10 Activity 1: Hematocrit Determination. Follow directions in the lab manual or from the handout at the front desk. 1. Record Data in the Table below: Blood Sample Height of Column of Blood (mm) Height of Red Blood Cell Layer (mm) Height of Buffy Coat (WBCs) (mm) Hematocrit %WBC Healthy male Boston Healthy female Boston Healthy male Denver Healthy female Denver Male aplastic anemia Female iron deficiency anemia 2. What is the hematocrit of a healthy male living at sea level in Boston? _______ 3. What is the hematocrit of a healthy female living at sea level in Boston? ______ 4. What is the hematocrit of a healthy male living at 1-mile elevation in Denver? ______ 5. How can you explain the difference between the hematocrits of the healthy male in Boston and the healthy male in Denver? Hint: look up the relationship of altitude to barometric pressure. Air with lower barometric pressure contains less oxygen. 6. What kidney hormone is released in response to low O2 levels? _________________ What is its target tissue? ______________________ What is its function? Activity 5: Total Cholesterol Determination Record Data in the Table Below Note: dL refers to deciliter (0.1 liter or 100 ml) Patient Approx. Total Cholesterol mg/dL Cholesterol Level 1 2 3 4 1. What are some essential roles for cholesterol in the body? 2. What risks are associated with low blood cholesterol? 3. What health problems might be in store for patient 2 based on these results? 4. What advice about diet and exercise would you give patient 4?