Instructions
When you have completed this assignment, you should send the results to Michael Ormerod by email.
Your answers need only be brief; there is no need to write an essay.
The written answers should be returned as a Word file.
If you areasked to send a picture, you should save it as a JPEG file.
When you have completed the assignment satisfactorily, you will have completed the Course. Congratulations!
There are four questions
Assignment
Question 1
In your laboratory, you have a wide range of instruments, including a conventional light microscope, a fluorescent microscope, a confocal microscope and a flow cytometer.
Two colleagues ask you for help with their measurements.
The first is working with a cell line grown in suspension culture. She wants to measure the percentage of cells expressing the intracellular protein, cyclin B. She also wants to measure whether the protein is located in the nucleus or the cytoplasm. Antibodies to cyclin B are available commercially.
The second is studying ovarian carcinoma. He wants to measure the percentage of ovarian carcinoma cells in peritoneal aspirates, in which cells are found in suspension. He also has biopsies of the solid tumours. He would like to know the distribution of infiltrating T lymphocytes within the tumour. He has antibodies to the ovarian tumour cells and to T lymphocytes, both are to antigens carried on the cell surface.
What is your advice to these colleagues? Give brief reasons.
Question 2
You are about to measure the number of B lymphocytes in a sample using a FITC-labelled monoclonal antibody to CD19.
Apart from green fluorescence, which other two parameters might you measure?
Which amplifier will you use to measure the green fluorescence – logarithmic or linear?
On which parameter will you set the threshold or discriminator setting?
Give brief reasons to justify your answers.