Garvan MLC flow Cytometry Facility guide to using Facs DiVa

1. MLC Flow Cytometry Facility
BD FACS DiVa
Training
Flow Cytometry

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. DiVa Overview

4. DiVa Overview
Browser
Window

5. DiVa Overview
Cytometer
Window
Browser
Window

6. DiVa Overview
Cytometer
Window
Worksheet
Browser Window
Window

7. DiVa Overview
Cytometer
Window
Worksheet
Browser Window
Window
Acquisition
Dashboard

8. DiVa Overview
Cytometer
Window
Worksheet
Browser Window
Window
Inspector Acquisition
Dashboard

9. DiVa Overview
Cytometer
Window
Worksheet
Browser Window
Window
Inspector Acquisition
Dashboard

10. File menu DiVa Overview
Cytometer
Window
Worksheet
Browser Window
Window
Inspector Acquisition
Dashboard

11. DiVa Windows -Browser
Organises
data in a
hierarchical
and ordered
manner

12. DiVa Windows - Browser

13. DiVa Windows - Browser

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. DiVa Windows - Cytometer

16. DiVa Windows -Cytometer
Parameter
name

17. DiVa Windows - Cytometer

18. DiVa Windows - Cytometer

19. DiVa Windows - Cytometer

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. Worksheets

22. Visualising Signal Data
Dot Density
plot plot
Statistical
measures are
also used to
Contour
plot identify
Histogram
changes

23. DiVa Windows - Acquisition
Dashboard

24. DiVa Windows - Acquisition
Dashboard

25. DiVa Windows - Acquisition
Dashboard

26. DiVa Windows - Acquisition
Dashboard

27. DiVa Windows – Inspector

28. DiVa Windows – Inspector

29. DiVa Windows – Inspector

30. DiVa Windows – Inspector

31. Creating a New Experiment

32. Creating a New Experiment

33. Creating a New Experiment

34. Compensation Workflow

35. Compensation Workflow

36. Compensation Workflow

37. Using the Layout editor

38. Using the Layout editor

39. Using the Layout editor

40. Fluidics Startup/Shutdown
Canto I, Canto II and Aria only

41. Fluidics Startup/Shutdown
Canto I, Canto II and Aria only

42. Fluidics Startup/Shutdown
Canto I, Canto II and Aria only

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. Exporting Data

45. Exporting Data

46. Exporting Data

47. Cleaning Instrument
1. Run 10 % fresh bleach soln. – 5 mins*
2. Run 4 % extran – 5 mins
3. Run distilled water – 5 mins
4. Shutdown if no other users in the day
* For Biohazardous samples

48. Troubleshooting

49. Troubleshooting