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Dear Prof. Smith and other experts Thanks for the replies and... sorry for bothering you again. As for the statistical model of two sample unpaired t-test with 2 nuisance variables (age, sex) in TBSS FA analysis... By using GLM GUI in FSL 4.1.4, I have set up the following design matrix (pls see the attached figures). And the 'design.con' file includes; /ContrastName1 control > patient /ContrastName2 patient > control /NumWaves 4 /NumContrasts 2 /PPheights 1.067873e+00 1.067873e+00 /RequiredEffect 1.617 1.617 /Matrix 1.000000e+00 -1.000000e+00 0.000000e+00 0.000000e+00 -1.000000e+00 1.000000e+00 0.000000e+00 0.000000e+00 I want to be sure that I am right, or please let me know the correct way. Thank you in advance for your kindness. Kim