2. What cells are in there? How can we use Flow Cytometry to quantify cells?
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6. How could you separate your sample of white blood cells into subpopulations? What characteristics of the cell could you use? How about if they had no color?
9. Flow Cytometry How it works 1. Draw cells, with excess fluid, from test tube into machine. 2. Cells pass in single file past laser. 3. Laser hits cell and light is scattered. 4. Photomultiplier multiplies light intensity and a light sensor measures the amount of light and scatter pattern. 5. Based on cell characteristics (size and shape), the computer categorizes and quantifies the cells. Click to learn more
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11. Use the following key to categorize your sample of White Blood Cells. What is the percent break up of your white blood cell differential? Action Lymphocyte White Nerds Eosinophil Pink Runt Basophil Yellow Runt Monocyte Green Runt Neutrophil Orange Runt Cell Candy
12. What can this differential tell you? http://www. bioteach . ubc .ca/ MolecularBiology / FlowCytometry / See real Data
13. Let’s take a closer look. How can we differentiate T cells? http://www.flow- cytometry .de/ Fluorescent Markers
14. Create a table to categorize and count your T-cell subpopulation into CD4+ Helper T cells and CD8+ Cytotoxic T cells. Purple Nerds= CD8+ Cytotoxic T cells Pink Nerds += CD4+ Helper T cells Categorize and count your T cell population Action
16. Homework What other applications are there for the technology of flow cytometry? Use the internet to explain 1 other application of the flow cytometer. E sure to document your resource.