Facs DiVa training Garvan MLC Flow Cytometry Facility

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Garvan MLC flow Cytometry Facility guide to using Facs DiVa

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Facs DiVa training Garvan MLC Flow Cytometry Facility

  1. 1. MLC Flow Cytometry FacilityBD FACS DiVaTraining Flow Cytometry
  2. 2. Login• Windows Login • User - Administrator • Password – BDIS or bdis• DiVa Login • Username - 3 letters first name + 3 letters last name • Password – as user name YOUR DISCIPLINE HERE
  3. 3. DiVa Overview
  4. 4. DiVa OverviewBrowserWindow
  5. 5. DiVa Overview Cytometer WindowBrowserWindow
  6. 6. DiVa Overview Cytometer Window WorksheetBrowser WindowWindow
  7. 7. DiVa Overview Cytometer Window WorksheetBrowser WindowWindow Acquisition Dashboard
  8. 8. DiVa Overview Cytometer Window WorksheetBrowser WindowWindow Inspector Acquisition Dashboard
  9. 9. DiVa Overview Cytometer Window WorksheetBrowser WindowWindow Inspector Acquisition Dashboard
  10. 10. File menu DiVa Overview Cytometer Window WorksheetBrowser WindowWindow Inspector Acquisition Dashboard
  11. 11. DiVa Windows -Browser Organises data in a hierarchical and ordered manner
  12. 12. DiVa Windows - Browser
  13. 13. DiVa Windows - Browser
  14. 14. DiVa Windows - Cytometer Status - instrument errors ( turns red) Parameters – controls PMT amplification and signal collection Threshold – controls triggering Laser – Laser delays and scaling Compensation – acquisition defined compensation matrix Ratio - ???
  15. 15. DiVa Windows - Cytometer
  16. 16. DiVa Windows -CytometerParametername
  17. 17. DiVa Windows - Cytometer
  18. 18. DiVa Windows - Cytometer
  19. 19. DiVa Windows - Cytometer
  20. 20. Worksheets• Global worksheet • Is not linked to specific data source • Reloads data on tube selection • Used when batching • Different global worksheets can be linked to different specimens• Sheet • Is linked to data • Does not refresh upon tube selection• Compensation • Is linked to compensation tubes • Must be used to perform acquisition compensation
  21. 21. Worksheets
  22. 22. Visualising Signal Data Dot Density plot plot Statistical measures are also used toContourplot identify Histogram changes
  23. 23. DiVa Windows - Acquisition Dashboard
  24. 24. DiVa Windows - Acquisition Dashboard
  25. 25. DiVa Windows - Acquisition Dashboard
  26. 26. DiVa Windows - Acquisition Dashboard
  27. 27. DiVa Windows – Inspector
  28. 28. DiVa Windows – Inspector
  29. 29. DiVa Windows – Inspector
  30. 30. DiVa Windows – Inspector
  31. 31. Creating a New Experiment
  32. 32. Creating a New Experiment
  33. 33. Creating a New Experiment
  34. 34. Compensation Workflow
  35. 35. Compensation Workflow
  36. 36. Compensation Workflow
  37. 37. Using the Layout editor
  38. 38. Using the Layout editor
  39. 39. Using the Layout editor
  40. 40. Fluidics Startup/Shutdown Canto I, Canto II and Aria only
  41. 41. Fluidics Startup/Shutdown Canto I, Canto II and Aria only
  42. 42. Fluidics Startup/Shutdown Canto I, Canto II and Aria only
  43. 43. Acquiring data• Activate tube to collect data• Hit acquire• Check stopping gate and numbers• Hit record• Hit remove tube• Remove tube and flick aspirator arm to left• repeate
  44. 44. Exporting Data
  45. 45. Exporting Data
  46. 46. Exporting Data
  47. 47. Cleaning Instrument1. Run 10 % fresh bleach soln. – 5 mins*2. Run 4 % extran – 5 mins3. Run distilled water – 5 mins4. Shutdown if no other users in the day * For Biohazardous samples
  48. 48. Troubleshooting
  49. 49. Troubleshooting

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