Introduce Flow cytometry, what are the major principles underlying this technique. With particular reference to this practical, discuss how you might use this technique to analyze the representation of various different hematopoietic subsets in the thymus and spleen
What’s CD3? If CD3 is positive what does it tell us about the cell?
What are the events that happen in the spleen and thymus that relate to T cell development/activation?
Insert a figure demonstrating how the assay works. (The figure legend should be well explained and it does not count towards the final word count)
(maximum 300 words)
Whats the aim of the practical? (doesn’t include the number of words)
Methods
Present an extremely brief overview of the methods used
DO NOT JUST COPY FROM THE METHOD THAT WAS GIVEN INSTEAD GIVE A BRIEF OVERVIEW AND EXPLAIN WHY YOU DID EACH STEP
(maximum 250 words)
Results
There is no maximum word count for any of the results section 1-5. HOWEVER, make sure that you discuss the data and the questions given.
Figure 1:
Present an annotated flow cytometry plot that best illustrates the percentage viable cells in the sample explaining why you might use these parameters to assess cell viability
Figure 2:
Present an annotated flow cytometry plot that demonstrates the percentage of CD4+ and CD8+ T cells in the spleen. What is the function of CD4+ and CD8+ T cells in the periphery?
Figure 3:
Present the two histograms that correlate with the presence of TCR on the T cell subsets described in Figure 2. Why can we use the presence of this marker on T cells to correlate with TCR expression?
Figure 4:
Present an annotated flow cytometry plot that demonstrates the development of CD4+ and CD8+ T cells in the thymus. What key events occur to the CD4+8+ (DP) thymocytes and how does affect the function of CD4+ and CD8+ T cells in the periphery?
Discussion
Discuss the relevance of your results presented in Figures 1 to 3 in terms of the immune activity that you would expect to happen in the spleen. What can we conclude from the results presented here about T cell development?
If you were using this technique to diagnose pathologies such as immunodeficiency, AIDS and leukemia, what would you expect to see in terms of T cell development and T cells in the periphery? Relate your answer to the flow cytometry data given.
