MLSP 300 Lab #4

Emely Peña Joydeep Mitra MLSP 300 Date of Lab Experiment: 3/28/24 Date of Lab Report Written: 4/5/24 Lab #4:Blood Analysis Introduction: In this lab we learned how to perform various types of blood analysis. These analyses include: hematocrit, leukocyte differential, manual red blood cell count, and manual white blood cell count. Hematocrit measures the proportion of red blood cells in the blood and % packed cell volume that aids in classifying anemia. Leukocyte differentiation measures the percent of each white blood cell type in the blood in a thin blood smear stained with biological dyes. Manual red blood cell count tells us the amount of red blood cells found in the blood; and manual white blood cell count tells us the amount of white blood cells found in the blood using a hemocytometer. We used the results from these tests for further calculations. Methods: In Part A of this lab, we performed a hematocrit test on the sample to measure the percent of packed cell volume. First, we filled three capillary tubes to about ¾ full with the blood sample

provided. Next, we gently pushed the capillary tubes into clay to seal the tubes off. Then, we placed our tubes into a centrifuge for five minutes and spinned it. Finally, we used a ruler and further calculations to determine the proportion of red blood cells. In Part B, we performed a leukocyte differential test to measure the percent of each white blood cell type in the blood sample provided. First, we filled two capillary tubes about ½ full with blood. Next, we added a drop of blood on two microscope slides for each of the capillary tubes and made a blood smear. Then, we stained the slides with biological dyes. After that, we placed the slides on a microscope and recorded the percentage of the different kinds of white blood cells found. Finally, using our observations/data and further calculations we determined the percentage of each white blood cell type in the blood sample. In Part C, we performed a manual red blood cell count to determine the amount of red blood cells found in the blood sample. First, we made a 1:200 dilution of the blood using 3.98mL of Gower’s solution as our diluent. Next, we mixed the dilution thoroughly. Then, we transferred about 20 μ L to a hemocytometer. After that, we counted the red blood cells in the five red blood cell grids and recorded our results. Finally, using our results and further calculation we found an estimated number of red blood cells in the original sample. In Part D, we performed a manual white blood cell count to determine the amount of white blood cells found in the blood sample. First, we made a 1:10 dilution of the blood using 0.90mL of Turk’s solution as our diluent. Next, we mixed the dilution thoroughly. Then, we transferred about 20 μ L to a hemocytometer. After that, we counted the white blood cells in the

Capillary Tube #2:

Capillary Tube #3:

#1 48.8fl #2 43.8fl #3 48.8fl Average 47.1fl Calculation for MCV: MCV = Hct * 10/RBC Count Capillary Tube #1: MCV = 29 * (10/ 5.94) MCV = 48.8 fl Capillary Tube #2: MCV = 26 * (10/ 5.94) MCV = 43.8 fl Capillary Tube #3: MCV = 29 * (10/ 5.94) MCV = 48.8 fl Average Capillary Tube MCV: MCV = 28 * (10/ 5.94) MCV = 47.1 fl Part B) Data Table: Leukocyte Differential

Type of Leukocyte Amount Counted Percentage Lymphocyte 10 45.0% Eosinophil 1 4.5% Neutrophil 9 41.0% Basophil 1 4.5% Monocyte 1 4.5% Part C) Data Table: Manual RBC Blood Count Square (1/25 sq. mm) Amount of RBC’s Inner upper Left 112 Inner upper Right 126 Center 116 Inner lower Left 117 Inner lower Right 123 Calculations: Average Amount of RBC per sq: 112 + 126 + 116 + 117 + 123 5 = 188.8 RBC’s per sq Total # RBC’s Counted = 594 RBC’s in 80/400 mm 2 Amount of RBC’s per liter = ( 594 0.02 mm 3 ) * ( 1 mm 3 1 μ L ) *200* 10 6 μ L 1 L = 5.94 *10 12 RBC’s per L

tubes was: 47.1fl. Our hematocrit and MCV are lower than the normal range can mean that the patient has anemia. In Part B, we found 5 leukocytes in our sample: lymphocyte (45%), eosinophil (4.5%), neutrophil (41%), basophil (4.5%), and monocyte (4.5%). We found lymphocytes, monocytes, basophils, eosinophils at slightly higher amounts and neutrophils in slightly lower amounts. We might’ve not observed/ counted correctly which could have caused these differences in amounts. In Part C, we found a manual red blood cell count of 5.94 *10 12 RBC’s per L. This amount is fine if the patient is male. In Part D, we found a manual white blood cell count of 1.56 *10 10 WBC’s per L. This amount is higher than the normal range. This may mean the patient has an infection that the immune system is trying to fight off. References: Lab Manual: Blood Analysis Questions and Problems: Report Findings For: 1. Hematocrit - Average: 28% - Capillary Tube #1: 29% - Capillary Tube #2: 26%

- Capillary Tube #3: 29% 2. Leukocyte Differential - Lymphocyte: 45% - Eosinophil: 4.5% - Neutrophil: 41% - Basophil: 4.5% - Monocyte: 4.5% 3. RBC Count - In 1 μ L : 5.94 *10 18 - In 1mL: 5.94 *10 15 - In 1L: 5.94 *10 12 4. WBC Count - In 1 μ L : 1.56 *10 16 - In 1mL: 1.56 *10 13 - In 1L: 1.56 *10 10