incubate with MTT and culture

6 Show the correct order for MTT assay to assess cell proliferation

a) Prepare cells, incubate with MTT and culture for 3 days and read at 550-600

b) Prepare cells, incubate with detergent, culture, add MTT, read at 550-600

c) Culture cells for 72 hours, add MTT, add detergent, read at 550-600

d) Culture with MTT and detergent for 72 hours, read at 550-600

cell counting using hemocytometer

4. Following cell isolation and cell counting using hemocytometer, you obtained 6X10 6 /ml ( 6 million

cells per milliliter). Total volume you have is 1 ml.

You are asked to set up cell cultures as follows: 1) control cells without any stimulant, 2) cells with

stimulant A, 3) cells with stimulant B, 4) cells with both stimulant A and B. Total four conditions.

You are asked to use 1.5X10 6 /ml per condition to set up the experiment.

Therefore, you have to dilute the cells you obtained after counting (6X10 6 /ml).

To prepare 1.5X10 6 /ml per condition, how much medium do you have to add to your 6X10

6 /ml.

EXPLAIN: SHOW THE WORK OUT DETAILS

cell concentration

3 peripheral blood lymphocyte cell samples were placed in hemocytomer and cells were counted in 3

quadrants. Total Cell counts in 3 quadrants were 80. Following final calculations cell concentration was

2.6X10 6 /ml. What is the dilution factor? Please show how you calculated?

a) Dilution factor is 5

b) Dilution factor is 2

c) Dilution factor if 10

d) Dilution factor is 20