Standard Operating Procedures s13

Purdue University Cytometry Laboratories Standard Operating Procedures

SOP-P057 How to Use ModfitLT Software

Objective: To use ModfitLT software for DNA analysis of flow cytometry listmode files

Procedure:

1. Access ModfitLT from the network by following SOP-C015. 2. Open Modfit by selecting the ModfitLT icon located on the desktop. 3. Click OK. 4. To open a listmode file for analysis click on either file>>open>> data, or click on the file button. 5. Go to the appropriate directory and folder where the files are located. 6. Click on the data file to be analyzed. 7. Click open 8. A Choose a parameter for analysis box will open. 9. select P5: FL4lin-FL4lin 10. click OK. 11. A Define Gates box will open. 12. click on Define Gate 1 (this will re-create the light scatter dot plot). 13. set parameter “x” to be P2: SSlin-SSlin set parameter “y” to be P1: FSlin-FSlin click OK 14. adjust the gate, but do not set the gate too tight. 15. click OK. 16. click on Define Gate 2 (this will re-create the doublet discrimination dot plot). 17. set parameter “x” to be P5: FL4lin-FL4lin 18. set parameter “y” to be P3: AUX-AUX 19. click OK. 20. adjust gate to gate out doublets. 21. click OK. 22. Will be returned to Define Gates box. 23. click OK. 24. select Mod (choose model) button from the top of the toolbar. 25. make selections based on the sample being analyzed. 26. click OK. 27. To move or adjust the ranges: left click and hold on the letter to move the region; left click and hold on the black square to change the size of the region. 28. click on the Fit button at the top of the toolbar. 29. If you want to add some text on the report, click on the Reprt (report) button at the top of the toolbar. A tools box will appear. Click on the “ A “ to open a text box. Type desired text and click on OK. 30. To print: select file>>print, should be set to pucl 8500 color printer. 31. To save the report: select file>>save as>>give name , will be *.mfl format. 32. When finished analyzing data, close ModfitLT and log out of the server by going to the bottom left of the desktop and clicking START>>Log Off verity>>YES.

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Created by: Cheryl Holdman Date: updated 8-29-06

Verified by: Date: Print Name Sign Name

of 1 Document Name: n:\manage\sops\sopP057.doc August 29, 2006