Flow cytometrypresentation

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Flow cytometrypresentation

  1. 1. Flow Cytometry TEACHER GUIDE SHEET Click to view POWERPOINT ACTIVITY
  2. 2. What cells are in there? How can we use Flow Cytometry to quantify cells?
  3. 3. <ul><li>What is flow Cytometry ? </li></ul><ul><li>Process for quantifying cells </li></ul><ul><ul><ul><li>Measures different property of cells </li></ul></ul></ul><ul><ul><ul><li>Able to categorize and quantify </li></ul></ul></ul><ul><ul><ul><li>Even able to separate out subpopulations of cells </li></ul></ul></ul>
  4. 4. Applications <ul><li>Clinical </li></ul><ul><li>Research </li></ul>Dr. Allan MCB Harvard
  5. 5. White Blood Cell Differentials <ul><ul><ul><li>Neutrophils </li></ul></ul></ul><ul><ul><ul><li>Lymphocytes </li></ul></ul></ul><ul><ul><ul><li>Monocytes </li></ul></ul></ul><ul><ul><ul><li>Eosinophils </li></ul></ul></ul><ul><ul><ul><li>Basophils </li></ul></ul></ul>White Blood Cells, Platelets (stained purple), a T-Lymphocyte white cell (stained green), and a Monocyte white cell (stained gold) as seen through a scanning electron microscope. ©2000 Dennis Kunkel, Ph.D. Image Source: http://www.mybloodyourblood.org/biology_white.htm
  6. 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?
  7. 7. Standard Differentials 1 2 3 <ul><li>Draw sample from patient </li></ul><ul><li>Smear blood on slide </li></ul><ul><li>Stain with differential WRIGHT stain </li></ul><ul><li>Count (at least 100) </li></ul>http://www. rnceus .com/ cbc / cbcwbc .html
  8. 8. Why do we need technology?
  9. 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
  10. 10. <ul><li>Create a table to record data for your white blood cell differential. </li></ul><ul><li>** Remember you will need to record the number of each cell in your sample. </li></ul><ul><ul><ul><li>Neutrophils </li></ul></ul></ul><ul><ul><ul><li>Lymphocytes </li></ul></ul></ul><ul><ul><ul><li>Monocytes </li></ul></ul></ul><ul><ul><ul><li>Eosinophils </li></ul></ul></ul><ul><ul><ul><li>Basophils </li></ul></ul></ul>Action
  11. 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. 12. What can this differential tell you? http://www. bioteach . ubc .ca/ MolecularBiology / FlowCytometry / See real Data
  13. 13. Let’s take a closer look. How can we differentiate T cells? http://www.flow- cytometry .de/ Fluorescent Markers
  14. 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
  15. 15. What can the results tell you? See real Data
  16. 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.
  17. 17. http://courses.dce.harvard.edu/-bios169c/resourcemain.html maintained by Dr. Jeffrey Lyczak Resources

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