Part 1:
1. Given the following dotplot of a cell preparation enriched for lymphocytes, draw what you would consider regions for:
- Platelets and debris
- Lymphocytes
- Monocytes/dendritic cells
- Granulocytes
2. Defend why each population distributes in such discrete areas on the dotplot:
- Platelets and debris-
- Lymphocytes-
- Monocytes/dendritic cells-
- Granulocytes-
3. If you were to gate on lymphocytes and stain with the following antibodies, draw in what you feel this dotplot would look like:
4. If you were to gate on lymphocytes and stain with the following antibodies, draw in what you feel this dotplot would look like:
5. If you stained all the cells (no regions or gates) with the following antibodies, draw in what you feel this dotplot would look like:
6. What is you stained just the lymphocytes with the following antibodies, predict what you feel the dotplot would look like:
Part 2:
Please open the file "final.851" from the zip "Flow Assignment Data"
Open this file in WinMDI and perform the following functions and answer the following questions. You do not have to attach the dotplot created in WinMDI, just enter the statistics requested and answer the questions associated.
The FITC label was anti-CD3, the PE label was anti-CD8 and the PerCP label was anti-CD19
First plot FSC vs. SSC, draw a region around the platelets/debris, lymphocytes, monocytes and granulocytes.
1. Report the percentage of each cell from the statistics of this step.
Plot FITC-CD3 vs. PE-CD8 and gate on the lymphocyte region.
2. Report what percentage of cells are in each quadrant and what type of cell you believe them to be (are they Th1, Th2, NK, what?).
3. How would you assess or test the function of the cells in the upper right quadrant?
Plot PE-CD8 vs. PerCP-CD19 and gate on the lymphocyte region.
4. Report what percentage of cells are in each quadrant and what type of cell you believe them to be (are they Th1, Th2, NK, what?).
5. How would you assess or test the function of the cells in the upper left quadrant?
Plot FITC-CD3 vs. PerCP-CD19 and gate on the lymphocyte region.
6. Report what percentage of cells are in each quadrant and what type of cell you believe them to be (are they Th1, Th2, NK, what?).
7. How would you assess or test the function of the cells in the lower left quadrant?
Attachment:- Flow Assignment Data.7z