ABO Blood Typing Lab Erin Kim 4B PART A
Introduction: In this lab, we tested blood (stimulated blood) to determine the ABO and the RH factor of four samples. The purpose of this was to basically understand blood transfusions and how they work. Materials:
4 Blood typing slides
12 Toothpicks
Unknown blood samples:
o Mr. Smith
o Mr. Jones
o Mr. Green
o Ms. Brown
Simulated Anti- A Serum
Simulated Anti- B Serum
Simulated Anti- Rh Serum
Procedure: Part A:
1. Label each blood typing slide: a. Slide 1: Mr. Smith
b. Slide 2: Mr. Jones c. Slide 3 Mr. Green
d. Slide 4: Ms. Brown 2. Place three to four drops of Mr. Smith’s blood in each of the A, B, and RH wells of Slide 1 3. Place three to four drops of Mr. Jones’s blood in each of the A, B, and RH wells of Slide 2 4. Place three to four drops of Mr. Green’s blood in each of the A, B, and RH wells of Slide 3 5. Place three to four drops of Ms. Brown’s blood in each of the A, B, and RH wells of Slide 4
a. The four labeled blood samples 6. Place three to four drops of simulated anti- A serum in each A well on the four slides
a. Adding the Anti- A serum to each A well.
7. Place three to four drops of simulated anti- B serum in each B well on the four slides 8. Place three to four drops of simulated anti- Rh serum in each Rh well on the four slides
a. Adding anti- RH serum to all the wells.
9. Obtain three toothpicks per blood typing slide. Stir each well with a separate clean toothpick for 30 seconds. To avoid splattering the simulated blood, do not press too hard on the typing try a. The stirring of the blood
10. Observe each slide and record your observation in Table 1 of the Analysis section. To confirm agglutination reaction.
11. Dispose of all materials according to your teacher’s instructions.
Data:
Table 1- Blood Typing
Anti-A Serum Anti-B Serum Anti-Rh Serum Blood Type Slide 1- Reaction No Reaction Reaction Type A Mr. Smith (Type A) (Positive) (Positive) Slide 2- No reaction Reaction No reaction Type B Mr. Jones (Type B) Negative Slide 3- Reaction No reaction Reaction Type A Mr. Green (Type A) (Positive) Positive Slide 4- No reaction No reaction No reaction Type O Ms. Brown Negative
Conclusion: The Data we saw was that Mr. Smith’s blood “clotted” in the A well and the RH well, but there was no reaction in the B well when the Anti- A, Anti-B, and the Anti- RH was put dropped in. Here we can see that Mr. Smith’s blood type is type A positive. The same steps were taken with Mr. Jones’, Mr. Green’s and Ms. Brown’s blood. Mr. Jones’s only reacted to the Anti B serum meaning that his blood type is Type B Negative. Mr. Green’s blood only had reactions to the Anti- A serum and the Anti- RH serum meaning that his blood was Type A Positive. Finally the last person was Ms. Brown unlike the three males, Ms. Brown had no reaction to any serums. This means she’s a Type O negative.
Evaluation: This lab was very successful, because we found out the blood types of all the people, but things we could have improved was the splatter of the blood. When we were mixing in the serums, our toothpicks flicked the blood everywhere, which made it possible for the different bloods/ serums to mix. Also instead of 12 toothpicks we ended up using 4 toothpicks, one for each blood type. This was a problem because the anti- a, b, and RH serums would be able to mix. Another problem was the timing of the mixing. When we didn’t see a reaction in 30 seconds of mixing, we tended to mix for another 10-20 seconds. In this lab we used simulated blood, so it wasn’t a big deal, but if we used real blood, and if we were in the real world it would be much more important to not make mistakes that we think are trivial in this lab. If we didn’t get their blood type right, then it could be extremely dangerous for them in blood transfusions. Part B
Introduction: After we determined the blood type of each blood sample, we took one sample, and made a wet mount slide. We placed it under the microscope to count each individual red blood cell and white blood cell. The purpose of counting each cell was to estimate the amount of white/ red cells in normal blood.
Materials
1 Microscope slide
1 Cover slip
Compound Microscope (400x magnification)
1 Blood sample. (Mr. Brown)
Procedure:
1. Thoroughly shake one of the vials of WARD’s Simulated Blood. Add one drop of simulated blood to a microscope slide, and cover with a cover slip. Lower the cover slip slowly to avoid trapping air bubbles on the slide
a. The blood sample slide we made.
2. Examine the slide with low power (10x). Find an area of the slide with an even distribution of cells. a. Compound Microscope
3. Switch to high power (40x). Refocus and count the number of simulated red blood cells (red spheres) in the field of view. Count the cells in any clump separately. Record the number in table 2 4. Count the number of simulated white blood cells (blue spheres). Record the number in table 2 5. Repeat the counting procedure with two other fields of view. Record these counts in Table 2 6. Calculate the average of the three red blood cell counts and the three white blood cell counts. Record the results in table 2 7. Multiply the average number of red and white blood cells by the dilution factor to determine the number of red and white blood cells per cubic millimeter. Record each value in Table 2
8. Dispose of all materials according to your teacher’s instructions
Table 2- Blood Cell Count.
Blood Cell Cell Count Total # Average # Dilution Total # Blood Cells Type of Cells of Cells Factor per mm3 or Avg. # Or Total/3 of Cells x Dilution 1 2 3 Factor Red (Red) 243 267 282 792 264 150,000 39,600,000 White 11 8 13 32 10.67 5,000 53,350 (Blue)
Conclusion:
In the second part of this lab, I learned so much about the numbers of red and white blood cells in your body. In this lab we counted every single cell in the square grid. We counted 3 of these grids, and averaged them out. We saw that the number of red cells was about 264 per grid, while the white blood cells were a lot smaller in number only have 10-11. By calculating the number of cells times the dilution factor I learned that the total number of blood cells per mm^3 was immense. There was 39600000 red blood cells and 53350 white blood cells. This lab helped me understand the importance of the white cells, and how there are so many more red cells than white cells.
Evaluation This part of the Lab was extremely difficult. Even though we had our microscope on 400x magnification, the cells we had to count were still extremely tiny. They were also moving all over the place, pressed under the cover slip. It was extremely hard to count for me, because I wear contacts and they were getting really dry. I had to blink every second and every time I blinked and opened my eyes the cells I was counting had moved, or I forgot where I had left off. We also had trouble seeing the white cells (blue). We couldn’t find many, and if we did we had trouble identifying if it was just a dust particle, or an actual cell. Our numbers are definitely off and could be off by even 100 cells. I would try to improve this next time by asking other people to help me count or I would bring eye drops! Even if that was the case, I’m pretty sure we couldn’t be 100% accurate.
This lab made me understand the Ratio of red cells to white cells. It kind of scared me that only that small number of white cells have to fight huge diseases such as cancers and tumors.