1. Human Anatomy and Physiology II Lab 2: Blood

2. Activity 1Blood Pathologies Observe prepared slides of blood smears taken from patients with the following disorders: pernicious anemia iron deficiency anemia sickle cell anemia chronic lymphocytic leukemia eosinophilia

3. Anemia refers to any condition in which there is a reduction in the number of RBCs or a reduction in the concentration of normal hemoglobin Anemia can be classified according to etiology (cause) or on the basis of morphology – For morphological classification, the following terms are used: RBC SizeRBC Color Microcytic: small sizeHypochromic: pale color Normocytic: normal sizeNormochormic: normal color Macrocytic: large sizeHyperchromic: dark color

4. Pernicous Anemia: caused by vitamin B 12 deficiency A.Can be either macrocytic, hyperchromic or macrocytic normochromic B.Note the enlarged, dark red blood cells, and the hypersegmented neutrophil in this smear

5. Iron Deficiency Anemia caused by iron insufficiency leading to decreased hemoglobin synthesis A.Is an example of microcytic, hypochromic anemia B.Note the small erythrocytes of varying sizes and the large area of central pallor

6. Sickle Cell Anemia: caused by a single amino acid substitution in the β chain of hemoglobin (valine replaces glutamic acid) A.This is a normochromic, normocytic anemia B.Note the sickle shaped erythrocytes.

7. Disorders Related to Leukocytes: I.Chronic Lymphocytic Leukemia caused by malignant lymphocyte progenitor cells in bone marrow A.Note the abundant small lymphocytes in this blood smear B.The lymphocyte nucleus may have a small indentation, or cleft

8. II.Eosinophilia many causes, including malignancy, connective tissue diseases, parasitic diseases, allergies; in some cases, no known cause Note the two normal looking eosinophils in the field

9. Activity 2 Blood Typing Pretested blood samples will be tested for the presence of A, B and Rh agglutinogens Type A Type B Type AB Type O Clinically Important Blood Groups Include the ABO System

10. And the Rh System – which includes the D agglutinogen Rh + Rh -

11. Blood Type is determined by determining which aggutinogens are present on RBC surfaces: Type A + Type AB + Type B - Type AB - Type O -

12. Agglutinins are antigens that combine with agglutinogens and cause a clumping reaction called agglutination + Type B CellsAnti-B agglutinin Agglutination

13. To determine the blood type of a sample, mix separate drops of a blood sample with anti A agglutinin, anti B agglutinin and anti-D agglutinin: Blood Typing Slide With 3 Drops Of Blood To Be Tested, One Drop In Each Well

14. Add Anti-A serum to this well and mix Add Anti-B serum to this well and mix Add anti-D serum to this well and mix Make sure that you use a separate toothpick for mixing in each well……………. If the agglutinin in the antiserum binds to agglutinogen you will soon observe agglutination

15. For example: Anti A Anti D Serum Serum Anti B Serum Type B + Type B - Type O +

16. This is what agglutination will look like !!!!!!!!!!!!!!!!!!!!!!!!

17. Safety Precautions for Dealing With Blood Samples: 1.Wear gloves 2.Wear goggles 3.Do not test your own blood – use our samples 4.Place any materials that contact blood in bleach solution immediately after use 5.Avoid spilling the samples – do not carry the slides or samples from place to place. If there is a spill, cover with bleach solution immediately and notify your lab instructor

18. Activity 3The ELISA Test The ELISA Test (Enzyme Linked ImmunoSorbant Assay): is an initial screening test for HIV, hepatitis B and hepatitis C infection In this activity, you will be test a serum sample from a fictional patient to see if they have a positive ELISA for anti-HIV antibody Individuals with healthy immune systems manufacture antibodies (anti-HIV antibodies) against HIV coat proteins, such as gp120

19. The HIV ELISA Screening Test is used to detect anti-HIV antibodies: Basic Steps: 1.A microplate with wells coated with HIV coat proteins (antigens) is used 2.A patients diluted serum is added to a well; if anti-HIV antibodies are present, they will specifically bind to the adherent coat proteins 3.A second antibody is added. This antibody is linked to an enzyme and it binds specifically to anti-HIV antibody. 4.A chromagen is added to the well; a chromagen is a colorless substrate that, when acted on by the enzyme, becomes colored (in this case, reddish yellow)

20. Each pair of students will be assigned a fictional patient (A – F) and will perform a test using the series of wells designated by the instructor Results need to be posted for class use ……..