Activity Owner : Jasmine Heng (Dr) Inputs By : Terence Chong (Dr), Ritu Bhalla (Dr) Approved By : Ventris Dsouza

(Dr) Module Chair : Ritu Bhalla (Dr)

A241: Cell culture P5 Practice questions : Counting down

1) John had trypsinized a flask of cells which were resuspended in a final volume

of 10 ml media. He used a 1:5 split ratio to start 2 new flasks. In addition, he took 1ml of the cells out (from the 10 ml of original cell suspension) and diluted it with an equal volume of trypan blue followed by cell count using a haemocytometer. Readings were taken from 10 primary squares and a total of 450 cells were counted.

(i)

What is the concentration of the cells in original suspension?

(ii)

What is the concentration of the cells in one of the new flask?

2) Andrew had pooled some kidney cells from 5 flasks together in a final volume

of 15ml of cell suspension. Using the haemocytometer, he counted the cells under the light microscope after adding an equal volume of trypan blue. He counted cells in 8 primary squares and obtained a final count of 600 cells. He needs to prepare 20ml of 1 X 105 cells/ml. How should he go about diluting the original cell suspension to get the required cell suspension?

3) Kim has prepared a cell suspension of 30ml of 3.5 X 10 6 cells/ml. What should

he do in order to prepare a sample stock of 7 X 106 cells/ml for his experiment?

4) Give an outline of a research problem where number of cells could be the

major tool for deriving conclusions of the study.