1. You are counting cells that you have prepared from a mouse spleen. Using the
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1. You are counting cells that you have prepared from a mouse spleen. Using the hemocytometer you determine that you have 5.8 x 106 cells/ml in your final suspension of cells. These cells are in a volume of 20 mls. Answer the following questions: a. How many total spleen cells did you obtain from this mouse (2 points)? b. You want to stain 106 cells with an antibody to CD4. How many milliters of the mouse spleen cell suspension do you need to use (2 points)?, c. The antibody to CD4 (anti-CD4) is available at a stock concentration of 500 ug/ml. When you stain the 106 cells from part "b", you will do so in a final volume of 0.1 ml, using an antibody concentration of 10 ug/ml. How much of the stock antibody do you need to add to your 0.1 ml solution to give you the appropriate final concentration (2 points)Explanation / Answer
a. 1 mL = 5.8 x 10^6 cells/mL
Total volume = 20 mL
Total number of cells = 20 X 5.8 x 10^6 cells
= 1.16 X 10^8 cells
b. 1 mL = 5.8 X 10^6 cells/mL
? mL = 1 X 10^6 cells
= (1 X 10^6)/(5.8 X 10^6)
= 0.1724 mL
= 172.4 uL
c. Antibody stock concentration = 500 ug/mL
Final volume = 0.1 mL
Final concentration = 10 ug/mL
Required stock concentration = (0.1 X 10)/500
= 1/500
= 0.002 mL
= 2 uL
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