ABO Blood Typing Lab Group members: Harri, Tracey, Emily, Arvind Oliver Yang Biology 4B Introduction: The purpose of this experiment is to identify the individual blood types. Based on the antigen/antibody reaction, we will be able to determine the individual blood types. Through using the Anti-­‐A, Anti-­‐B, and Anti-­‐Rh Serums, we will be able to ascertain the specific blood types of each individual. Hypothesis: Based on the antigen/antibody specific reaction, we can determine the ABO and Rh blood types using specific antiserum. This is due to the fact that if an A Antigen comes into contact with an Anti A Antibody, the blood will have a physical reaction. If a B Antigen comes into contact with an Anti B Antibody, the blood will also seem to condense in texture. Materials: 1) Four different simulated sample blood types (provided by Ward’s Simulated Blood) 2) Simulated Anti-­‐A Serum (Provided by Ward’s Natural Science) 3) Simulated Anti-­‐B Serum (Provided by Ward’s Natural Science) 4) Simulated Anti-­‐Rh Serum (Provided by Ward’s Natural Science) 5) Four Blood Typing slides (Provided by Ward’s Natural Science) 6) Compound Microscope 7) Microscope slide 8) Cover slip 9) Twelve Toothpicks Procedure: Part 1: 1) Label each different simulated Blood type a name, consisting of Mr. Smith, Mr. Jones, Mr. Green, and Ms. Brown. 2) On Slide #1, place 3-­‐4 drops of Mr. Smith’s blood in the A, B, and O wells 3) On Slide #2, place 3-­‐4 drops of Mr. Jones’ blood in the A, B, and O wells 4) On Slide #3, place 3-­‐4 drops of Mr. Green’s blood in the A, B, and O wells 5) On Slide #4, place 3-­‐4 drops of Ms. Brown’s blood in the A, B, and O wells 6) On each of the four slides, drop 3-­‐4 drops of Anti-­‐A Serum in each of the A wells located on each slide. 7) On each of the four slides, drop 3-­‐4 drops of Anti-­‐B Serum in each of the B wells located on each slide 8) On each of the four slides, drop 3-­‐4 drops of Anti-­‐Rh Serum in each of the Rh wells located on each slide 9) Use three toothpicks per slide, one toothpick per well. Maintain cleanliness of each toothpick by using one for EACH well. 10) Using the toothpicks, stir each well for 30 seconds 11)Observe the changes in each well after stirring each for 30 seconds. Part 2: 1) Shake one of the vials of simulated blood. 2) Take one microscope slide and add one drop of simulated blood onto the slide. 3) Cover the slide with a cover slip, try to not get any air bubbles underneath the cover slip. 4) Using 10x magnification, find an area in which there is an even distribution of cells (small spheres) 5) After locating one area, switch to 40x magnification and count the number of red blood cells (red spheres) in the area, and also the number of white blood cells (blue spheres) in the area. Record these numbers 6) Repeat Step 5 for a different area, record in a different table 7) Repeat Step 5 for another different area, record in a different table 8) Calculate the average and number of red and white blood cells per cubic millimeter Data: Table 1 (Part 1): Slide #1 Mr. Smith Slide #2 Mr. Jones Slide #3 Mr. Green Slide #4 Ms. Brown Anti-­‐A Anti-­‐B Serum Serum Reaction No Reaction (Type A) No Reaction Reaction (Type B) Reaction No reaction (Type A) No Reaction No Reaction Anti-­‐Rh Serum Reaction (positive) No Reaction (negative) Reaction (Positive) No Reaction Blood Type Type A Positive Type B Negative Type A Positive Type 0 Table 2 (Part 2): Blood Cell Count Cell 1 2 3 Type Red 300 321 302 White 7 8 7 Total number of Cells 923 22 Average Number cells (total/3) 308 7 Dilution Factor 150,000 5,000 Blood sample: Jones Picture #1: Dropping specific blood types into slides Total no. blood cells per mm3 (Avg. no. cells x dilution factor) 46,200,000 35,000 Picture #2: Stirring the slides that contain specific blood types Picture #3: Applying the antiserum to each of the blood types in their specific wells Picture #4: Looking through the microscope to count cells Conclusion: Through part 1 of this experiment we can conclude that Mr. Smith’s and Mr. Green’s blood type is Type A Positive. Mr. Jones has Type B Negative blood. Ms. Brown has Type O blood. All of these conclusions were reached by using specific antiserums with specific blood samples. In the case of Mr. Jones, we were able to notice a physical reaction after we placed 3-­‐4 drops of antiserum into the B well and stirring it. This was also apparent for Mr. Smith and Mr. Green, but only in the A wells. Ms. Brown had no reaction in any of the wells. The purpose of knowing this process is key to the evaluation of finding if someone is able to follow through a blood transfusion. Through part 2 of this experiment we are able to conclude that Mr. Jones’ blood has an average number of 308 red blood cells and 7 white blood cells in each quadrant. We were also able to calculate the total number of blood per cubic millimeter. The purpose of doing this kind of experiment is to help determine if someone has a blood-­‐related disease, like anemia. Evaluation: Although this experiment did give out plausible data, some factors have to be accounted for. While stirring with the toothpick I found myself pushing too hard on the slide. This created a lot of splattering, which may have caused some blood to go into other wells in its vicinity, causing some of our data to be off. Looking to improve this fault, I would suggest using a shaker machine. This would halt human error to a stop and also prevent splattering from occurring. Another weakness I possibly made during the course of the experiment was during the counting of red blood cells and white blood cells. Due to trying to finish in time, I might have skipped a couple of red blood cells. If we were to do this experiment again, I would make sure to take my time when counting.