You will perform a serial dilution in class.  This is a doubling dilution which halves the concentration of cells with each dilution.

At Step 4 in Part B you will create different ratios of effector cells to target cells.  What is the number of cells in wells A1, A4, A7, and A10 following a serial dilution?  (Answer in standard notation (e.g. 8000 rather than 8 × 10³.)

A1:

Which haplotype/s does each mouse strain express?

Based on this practical exercise, for the following conditions indicate if you would expect a high level of cell killing, an intermediate level of cell killing, or no cell killing.

You will perform a serial dilution in class.  This is a doubling dilution which halves the concentration of cells with each dilution.

At Step 4 in Part B you will create different ratios of effector cells to target cells.  What is the number of cells in wells A1, A4, A7, and A10 following a serial dilution?  (Answer in standard notation (e.g. 8000 rather than 8 × 10³.)

A1:

What is the purpose of the RPMI control in the cytotoxicity assay?

You have a cell suspension for which you need to determine the concentration of cells/ml.  You take 20 μl of cell suspension and this with 60 μl of counting solution.  You perform a haemocytometer count, and you have counted 62, 72, 61, and 68 cells in the four large outer squares (A, B, C, and D) of the haemocytometer.

What is the concentration (in cells/ml) of your sample? (Answer in standard notation (e.g. 62000 instead of 6.2 × 10⁴).)

You have 4 ml of a cell suspension with a concentration of 5.9 × 10⁵ cells/ml.  You need to make up 2 ml of cells at a concentration of 1 × 10⁶ cells/ml.  What volume of the original cell suspension will you need to make up 2 ml of cells at a 1 × 10⁶ cells/ml? ml. (Answer to 2 decimal places.)

As you need 2 mls of cells at the desired concentration you centrifuge and pallet your cells, and then remove the liquid from the pallet.  How much buffer do you need to add to these cells to make up 2 ml of cells at 1 ×10⁶ cells/ml? μl.

(Check the units carefully.  If you are not sure how to use C₁V₁ = C₂V₂ to answer this question refer to the information in the Numbers, graphs and calculations eBook.)

You've performed a haemocytometer count and have determined that there are 1.92 × 10⁸ cells/ml in your 2 ml of cell suspension.

In the MHC restriction of T cells experiment you will need to make up a 1 ml aliquot of 1 × 10⁶ cells.  You will need to remove μl of cell suspension and add μl of buffer to make up this cell suspension.

(Answer to 2 decimal places.)

Hint: If you are not sure how to complete this calculation check the C₁V₁ = C₂V₂ video in the Numbers, graphs and calculations eBook.

In the early stages of T cell development in the thymus, a thymocyte that has just completed its expression of a TCR will:

How many different MHC class I molecules does each nucleated cell in our body express?