You did an experiment that required to have a large number 5

You did an experiment that required to have a large number, 5,animals /group. At the day of the assay you pooled the spleens of each group. You made single cell suspension, washed the cells and reconstituted the spleens in 30 mls of media. Subsequently you diluted your stock 5 fold and took a small sample from this tube and you diluted it further 1:2 in Trypan Blue You counted viable white cells present in the spleen. You counted 105 cells in 15 squares of the 25 squares of the L5 region of the Hemocytometer. Use the stock cells of the above experiment and do the followings:

- Make serial two-fold dilutions. Calculate the concentration of cells/1 ml in the 6th dilution tube Indicate with volumes of your choice of how you can do 2 fold dilutions

- Calculate the Total number of cells present in 7th dilution tube (remember the volumes used for these dilutions are of your choice)

Solution

Answer:

Cells/uL = (cells x Dilution factor )/(number of squares x 0.1)

Dilution factor = 5 x 2 =10

Cells/uL = (105 x 10 )/(15 x 0.1)

Cells/uL = 700

Cells/ mL = 700 x 1000

Tube 1 = One mL of above stock of cells + One mL of diluent media = 2 fold dilution

Tube 2 = 4 fold dilution

Tube 3 = 8 fold dilution

Tube 4 = 16 fold dilution

Tube 5 = 32 fold dilution

Tube 6 = 64 fold dilution

concentration of cells/1 ml in the 6th dilution tube = (700 x 1000)/64 =10937.5 cells/mL