Our entry in the Functional Imaging Analysis contest Jonathan Taylor Stanford Keith Worsley McGill What is functional Magnetic Resonance Imaging (fMRI) data? Time series of ~200 “frames”, 3D images of brain “activity”, taken every ~2.5s (~8min) Meanwhile, subject receives stimulus or external task (e.g on/off every 10s) Several (~4) time series (“runs”) per session Several (~2) sessions per subject Several (~15) subjects Statistics problem: find the regions of the brain activated by the stimulus or task Why a Functional Imaging Analysis Contest (FIAC)? Competing packages produce slightly different results, which is “correct”? Simulated data? Real data, compare analyses “Contest” session at 2005 Human Brain Map conference 9 entrants Results in a special issue of Human Brain Mapping in May, 2006 The main participants SPM (Statistical Parametric Mapping, 1993), University College, London, “SAS”, (MATLAB) AFNI (1995), NIH, more display and manipulation, not much stats (C) FSL (2000), Oxford, the “upstart” (C) …. FMRISTAT (2001), McGill, stats only (MATLAB) BRAINSTAT (2005), Stanford/McGill, Python version of FMRISTAT Effect of stimulus on brain response Alternating hot and warm stimuli separated by rest (9 seconds each). 2 1 0 -1 0 50 100 150 200 250 300 350 Hemodynamic response function: difference of two gamma densities Stimulus is delayed and dispersed by ~6s 0.4 0.2 0 Modeled by convolving the stimulus with the “hemodynamic response function” -0.2 0 50 Responses = stimuli * HRF, sampled every 3 seconds 2 1 0 -1 0 50 100 150 200 Time, seconds 250 300 350 fMRI data, pain experiment, one slice First scan of fMRI data Highly significant effect, T=6.59 1000 hot rest warm 890 880 870 500 0 100 200 300 No significant effect, T=-0.74 820 hot rest warm 0 800 T statistic for hot - warm effect 5 0 -5 T = (hot – warm effect) / S.d. ~ t110 if no effect 0 100 0 100 200 Drift 300 810 800 790 200 Time, seconds 300 How fMRI differs from other repeated measures data Many reps (~200 time points) Few subjects (~15) Df within subjects is high, so not worth pooling sd across subjects Df between subjects low, so use spatial smoothing to boost df Data sets are huge ~4GB, not easy to use R directly FMRISTAT / BRAINSTAT statistical analysis strategy Analyse each voxel separately Break up analysis into stages Borrow strength from neighbours when needed 1st level: analyse each time series separately 2nd level: combine 1st level results over runs 3rd level: combine 2nd level results over subjects Cut corners: do a reasonable analysis in a reasonable time (or else no one will use it!) MATLAB / Python 1st level: Linear model with AR(p) errors Data Model Yt = fMRI data at time t xt = (responses,1, t, t2, t3, … )’ to allow for drift Yt = xt’β + εt εt = a1εt-1 + … + apεt-p + σFηt, ηt ~ N(0,1) i.i.d. Fit in 2 stages: 1st pass: fit by least squares, find residuals, estimate AR parameters a1 … ap 2nd pass: whiten data, re-fit by least squares Higher levels: Mixed effects model Data Model Ei = effect (contrast in β) from previous level Si = sd of effect from previous level zi = (1, treatment, group, gender, …)’ Ei = zi’γ + SiεiF + σRεiR (Si high df, so assumed fixed) εiF ~ N(0,1) i.i.d. fixed effects error εiR ~ N(0,1) i.i.d. random effects error Fit by ReML Use EM for stability, 10 iterations Where we use spatial information 1st level: smooth AR parameters to lower variability and increase “df” Higher levels: smooth Random / Fixed effects sd ratio to lower variability and increase “df” Final level: use random field theory to correct for multiple comparisons 1st level: Autocorrelation AR(1) model: εt = a1 εt-1 + σFηt Fit the linear model using least squares εt = Y t – Y t â1 = Correlation (εt , εt-1) Estimating errort’s changes their correlation structure slightly, so â1 is slightly biased: Raw autocorrelation Smoothed 12.4mm ~ -0.05 Bias corrected â1 ~0 0.3 0.2 0.1 0 -0.1 How much smoothing? • Variability in acor lowers df • Df depends on contrast • Smoothing acor brings df back up: ( dfacor = dfresidual 1 1 dfeff = dfresidual+ Hot stimulus FWHMacor2 2 +1 2 FWHMdata 2 acor(contrast of data)2 dfacor FWHMdata = 8.79 Residual df = 110 0 100 Target = 100 df Contrast of data, acor = 0.61 dfeff 0 10 20 FWHM = 10.3mm Hot-warm stimulus Residual df = 110 100 Target = 100 df 50 ) 3/2 Contrast of data, acor = 0.79 50 dfeff 30 FWHMacor 0 0 10 20 FWHM = 12.4mm 30 FWHMacor Higher order AR model? Try AR(3): a1 a2 a3 0.3 0.2 AR(1) seems to be adequate 0.1 0 … has little effect on the T statistics: No correlation AR(1), df=100 AR(2), df=99 -0.1 AR(3), df=98 5 0 -5 biases T up ~12% → more false positives 2nd level: 4 runs, 3 df for random effects sd Run 1 Run 2 Run 3 Run 4 2nd level Effect, Ei 1 0 … very noisy sd: -1 0.2 Sd, Si 0.1 … and T>15.96 for P<0.05 (corrected): 0 5 T stat, E i / Si 0 … so no response is detected … -5 Solution: Spatial smoothing of the sd ratio • Basic idea: increase df by spatial smoothing (local pooling) of the sd. • Can’t smooth the random effects sd directly, - too much anatomical structure. • Instead, sd = smooth random effects sd fixed effects sd fixed effects sd ) which removes the anatomical structure before smoothing. ^ Average Si Random effects sd, 3 df Fixed effects sd, 440 df Mixed effects sd, ~100 df 0.2 0.15 0.1 0.05 divide Random sd / fixed sd 0 multiply Smoothed sd ratio 1.5 1 0.5 random effect, sd ratio ~1.3 How much smoothing? ( dfratio = dfrandom FWHMratio2 2 +1 2 FWHMdata ) 1 1 1 = + dfeff dfratio dffixed 3/2 dfrandom = 3, dffixed = 4 110 = 440, FWHMdata = 8mm: fixed effects analysis, dfeff = 440 400 300 dfeff Target = 100 df random effects analysis, dfeff = 3 200 FWHM = 19mm 100 0 0 20 40 FWHMratio Infinity Final result: 19mm smoothing, 100 df Run 1 Run 2 Run 3 Run 4 2nd level Effect, Ei 1 0 … less noisy sd: -1 0.2 Sd, Si 0.1 … and T>4.93 for P<0.05 (corrected): 0 5 T stat, E i / Si 0 … and now we can detect a response! -5 Final level: Multiple comparisons correction Bonferroni 4.7 4.6 Gaussianized threshold 4.5 True 4.4 4.3 4.2 T, 10 df Random Field Theory T, 20 df Discrete Local Maxima (DLM) 4.1 Gaussian 4 3.9 3.8 3.7 0 Low FWHM: use Bonferroni 1 2 3 4 5 6 7 8 FWHM of smoothing kernel (voxels) In between: use Discrete Local Maxima (DLM) 9 10 High FWHM: use Random Field Theory 0.12 Gaussian T, 20 df T, 10 df 0.1 Random Field Theory Bonferroni P-value 0.08 0.06 0.04 True DLM can ½ P-value when FWHM ~3 voxels 0.02 0 Discrete Local Maxima 0 Low FWHM: use Bonferroni 1 2 3 4 5 6 7 FWHM of smoothing kernel (voxels) In between: use Discrete Local Maxima (DLM) 8 9 10 High FWHM: use Random Field Theory FIAC paradigm 16 subjects 4 runs per subject 4 conditions per run 2 runs: event design 2 runs: block design Same sentence, same speaker Same sentence, different speaker Different sentence, same speaker Different sentence, different speaker 3T, 191 frames, TR=2.5s Response Events 0.4 0.2 0 -0.2 0 50 100 150 200 250 300 350 400 450 500 350 400 450 500 Beginning of block/run Blocks 0.4 0.2 0 -0.2 0 50 100 150 200 250 Seconds 300 Design matrix for block expt B1, B2 are basis functions for magnitude and delay: 1st snt in block S snt, S spk, B1 S snt, S spk, B2 S snt, D spk, B1 S snt, D spk, B2 D snt, S spk, B1 D snt, S spk, B2 D snt, D spk, B1 D snt, D spk, B2 Constant Linear Quadratic Cubic Spline Whole brain avg 1st level analysis Motion and slice time correction (using FSL) 5 conditions 3 contrasts Beginning of block/run Same sent, Same sent, same speak diff speak Diff sent, same speak Diff sent, diff speak Sentence Speaker 0 0 -0.5 -0.5 -0.5 0.5 0.5 -0.5 0.5 0.5 1 -1 -1 1 Interaction 0 Smoothing of temporal autocorrelation to control the effective df Magnitude sd (relative to error) Efficiency 2 1.5 Sd of contrasts (lower is better) for a single run, assuming additivity of responses Event Block 1 0.5 0 Diff sente Diff speak • For the magnitudes, event and block have similar efficiency • For the delays, event is much better. Interac Delay sd (seconds) 1.6 1.4 1.2 1 0.8 0.6 0.4 0.2 0 Event Block Diff sente Diff speak Interac 2nd level analysis Analyse events and blocks separately Register contrasts to Talairach (using FSL) Bad registration on 2 subjects - dropped Combine 2 runs using fixed FX 3rd level analysis Combine remaining 14 subjects using random FX 3 contrasts × event/block × magnitude/delay = 12 Threshold using best of Bonferroni, random field theory, and discrete local maxima (new!) Part of slice z = -2 mm Magnitude (%BOLD), diff - same sentence, event experiment Subj 0 1 3 4 6 7 8 9 10 11 12 13 14 Mixed effects 15 Left 2 1 0 Ef Right -1 Random /fixed effects sd smoothed 7.0105mm 1.5 -2 Slice range is -74<x<70mm, -46<y<4mm, z=-2mm; Contour is: min fMRI > 6214 1 Left Sd Right 271 272 271 265 264 132 270 275 269 274 248 256 264 278 1 0 0.5 5 FWHM (mm) 15 40 Left -50 T 0 x (mm) df 0.5 10 0 5 50 Ant. Post. Right P=0.05 threshold for local maxima is +/- 5.68 -5 -40-20 0 y (mm) 0 Magnitude (%BOLD), diff - same sentence, block experiment Subj 0 1 3 4 6 7 8 9 10 11 12 13 14 Mixed effects 15 Left 2 1 0 Ef Right -1 Random /fixed effects sd smoothed 7.103mm 1.5 -2 Slice range is -74<x<70mm, -46<y<4mm, z=-2mm; Contour is: min fMRI > 5904 1 Left Sd Right 202 202 204 205 204 203 201 202 200 206 205 202 204 200 1 0 0.5 5 FWHM (mm) 15 40 Left -50 T 0 x (mm) df 0.5 10 0 5 50 Ant. Post. Right P=0.05 threshold for local maxima is +/- 5.67 -5 -40-20 0 y (mm) 0 Delay shift (secs), diff - same sentence, event experiment Subj 0 1 3 4 6 7 8 9 10 11 12 13 14 Mixed effects 15 Left 0.2 0.1 0 -0.1 -0.2 Ef Right Random /fixed effects sd smoothed 10.6778mm 1.5 Slice range is -74<x<70mm, -46<y<4mm, z=-2mm; Contour is: magnitude, stimulus average, T statistic > 5 Left 0.4 Sd 1 0.2 Right df 271 272 271 265 264 132 270 275 269 274 248 256 264 278 0 0.5 40 FWHM (mm) 15 T 0 -2 Ant. Post. Right P=0.05 threshold for local maxima is +/- 4.31 -50 x (mm) Left 2 10 0 5 50 -40-20 0 y (mm) 0 Delay shift (secs), diff - same sentence, block experiment Subj 0 1 3 4 6 7 8 9 10 11 12 13 14 Mixed effects 15 Left 1 0.5 0 Ef Right -0.5 Random /fixed effects sd smoothed 8.8952mm 1.5 -1 Slice range is -74<x<70mm, -46<y<4mm, z=-2mm; Contour is: magnitude, stimulus average, T statistic > 5 2 Left 1.5 Sd 1 1 0.5 Right df 202 202 204 205 204 203 201 202 200 206 205 202 204 200 0 0.5 40 FWHM (mm) 15 T 0 -2 Ant. Post. Right P=0.05 threshold for local maxima is +/- 4.3 -50 x (mm) Left 2 10 0 5 50 -40-20 0 y (mm) 0 Event Block Subj 0 1 3 4 6 7 8 9 10 11 12 13 14 Magnitude (%BOLD), diff - same sentence, block experiment Mixed effects 15 1 3 4 6 7 8 9 10 11 12 13 14 Left Left 0 0 Random /fixed effects sd smoothed 7.0105mm 1.5 -2 1 1 Left 271 265 264 132 270 275 269 274 248 256 264 278 Sd 0.5 1 0 0.5 40 Right Right 272 Random /fixed effects sd smoothed 7.103mm 1.5 -2 Slice range is -74<x<70mm, -46<y<4mm, z=-2mm; Contour is: min fMRI > 5904 Left Sd 271 -1 Right Right -1 2 1 Ef Slice range is -74<x<70mm, -46<y<4mm, z=-2mm; Contour is: min fMRI > 6214 df Mixed effects 15 1 Ef Magnitude Subj 0 2 df 202 202 204 205 204 203 201 202 200 206 205 202 204 200 x (mm) 10 FWHM (mm) 15 -50 T 0 0.5 Left Left 0 0 5 -50 T 1 40 FWHM (mm) 15 5 0.5 0 5 x (mm) Magnitude (%BOLD), diff - same sentence, event experiment 50 1 3 4 6 7 8 9 10 11 12 13 14 -40-20 0 y (mm) P=0.05 threshold for local maxima is +/- 5.67 Delay shift (secs), diff - same sentence, block experiment Mixed effects 15 Subj 0 1 3 4 6 7 8 9 10 11 12 13 14 Mixed effects 15 Left Left 0.2 0.1 0 -0.1 -0.2 Ef -0.5 2 1.5 Sd 1 0.2 271 272 271 265 264 132 270 275 269 274 248 256 264 278 1 0.5 0 Right Right df Random /fixed effects sd smoothed 8.8952mm 1.5 -1 Left Left 1 0.5 40 df 202 202 204 205 204 203 201 202 200 206 205 202 204 200 0 0.5 40 -2 10 T 0 2 -50 0 5 50 0 P=0.05 threshold for local maxima is +/- 4.3 Ant. -40-20 0 y (mm) -2 Post. Right Ant. Post. Right P=0.05 threshold for local maxima is +/- 4.31 x (mm) 0 -50 Left Left T FWHM (mm) 15 x (mm) FWHM (mm) 15 2 0 1 Slice range is -74<x<70mm, -46<y<4mm, z=-2mm; Contour is: magnitude, stimulus average, T statistic > 5 0.4 Sd -40-20 0 y (mm) 0 Right Right Random /fixed effects sd smoothed 10.6778mm 1.5 -5 0.5 Ef Slice range is -74<x<70mm, -46<y<4mm, z=-2mm; Contour is: magnitude, stimulus average, T statistic > 5 Delay 0 Ant. Subj 0 5 50 Post. Right Delay shift (secs), diff - same sentence, event experiment -5 Ant. Post. Right P=0.05 threshold for local maxima is +/- 5.68 10 0 10 0 5 50 -40-20 0 y (mm) 0 Events vs blocks for delays in different – same sentence Events: 0.14±0.04s; Blocks: 1.19±0.23s Both significant, P<0.05 (corrected) (!?!) Answer: take a look at blocks: Greater magnitude Best fitting block Greater delay Different sentence (sustained interest) Same sentence (lose interest) SPM BRAINSTAT Magnitude increase for Sentence, Event Sentence, Block Sentence, Combined Speaker, Combined at (-54,-14,-2) Magnitude decrease for Sentence, Block Sentence, Combined at (-54,-54,40) Delay increase for Sentence, Event at (58,-18,2) inside the region where all conditions are activated Conclusions Greater %BOLD response for different – same sentences (1.08±0.16%) different – same speaker (0.47±0.0.8%) Greater latency for different – same sentences (0.148±0.035 secs) The main effects of sentence repetition (in red) and of speaker repetition (in blue). 1: Meriaux et al, Madic; 2: Goebel et al, Brain voyager; 3: Beckman et al, FSL; 4: Dehaene-Lambertz et al, SPM2. z=-12 z=2 2 3 z=5 1 1,4 3 Brainstat: combined block and event, threshold at T>5.67, P<0.05. 3 3 1 3 Estimating the delay of the response • Delay or latency to the peak of the HRF is approximated by a linear combination of two optimally chosen basis functions: delay 0.6 0.4 basis1 0.2 HRF basis2 0 -0.2 -0.4 -5 0 shift 5 10 t (seconds) 15 20 25 HRF(t + shift) ~ basis1(t) w1(shift) + basis2(t) w2(shift) • Convolve bases with the stimulus, then add to the linear model • Fit linear model, estimate w1 and w2 3 w2 / w1 2 1 • Equate w2 / w1 to estimates, then solve for shift (Hensen et al., 2002) w1 • To reduce bias when the magnitude is small, use 0 w2 shift / (1 + 1/T2) -1 where T = w1 / Sd(w1) is the T statistic for the magnitude -2 -3 -5 0 shift (seconds) 5 • Shrinks shift to 0 where there is little evidence for a response. Magnitude (%BOLD), stimulus average 0 1 3 4 6 7 Subject id, event experiment 8 9 10 11 12 13 14 15 Mixed effects 6 4 Ef 2 0 Random /fixed effects sd smoothed 10.836mm -2 Contour is: average anatomy > 2000 2 1.5 1.5 Sd 1 1 0.5 200 200 200 200 200 100 200 200 200 200 200 200 200 200 0.5 5 FWHM (mm) 20 50 15 100 10 150 100 T 0 x (mm) df 0 5 200 -5 P=0.05 threshold for peaks is +/- 5.1687 250 120 140 160 180 y (mm) 0 Magnitude (%BOLD), stimulus average 0 1 3 4 6 7 Subject id, block experiment 8 9 10 11 12 13 14 15 Mixed effects 4 Ef 2 Random /fixed effects sd smoothed 6.7765mm 0 Contour is: average anatomy > 2000 1 6 Sd 0.5 4 2 0 df 209 209 214 210 211 210 210 207 212 214 214 210 210 216 99 FWHM (mm) 20 20 -50 T 10 5 0 -5 P=0.05 threshold for peaks is +/- 5.9873 x (mm) 15 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Magnitude (%BOLD), diff - same speaker 0 1 3 4 6 7 Subject id, event experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 11.7848mm 1.5 -2 Contour is: average anatomy > 2000 2 1.5 Sd 1 1 0.5 df 273 271 276 281 274 136 265 293 264 268 265 264 296 270 0 0.5 5 FWHM (mm) 20 100 T 0 -5 P=0.05 threshold for peaks is +/- 5.1469 x (mm) -50 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Magnitude (%BOLD), diff - same speaker 0 1 3 4 6 7 Subject id, block experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 11.9735mm 1.5 -2 Contour is: average anatomy > 2000 1 Sd df 201 202 200 206 201 201 200 200 204 204 206 201 205 204 0.5 1 0 0.5 5 FWHM (mm) 20 100 T 0 -5 P=0.05 threshold for peaks is +/- 5.1666 x (mm) -50 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Magnitude (%BOLD), interaction 0 1 3 4 6 7 Subject id, event experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 11.4737mm 1.5 -2 Contour is: average anatomy > 2000 2 1.5 Sd 1 1 0.5 df 278 278 279 280 264 126 277 287 264 272 260 277 264 264 0 0.5 5 FWHM (mm) 20 100 T 0 -5 P=0.05 threshold for peaks is +/- 5.4124 x (mm) -50 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Magnitude (%BOLD), interaction 0 1 3 4 6 7 Subject id, block experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 12.1993mm 1.5 -2 Contour is: average anatomy > 2000 1 Sd df 204 200 207 200 204 205 202 203 202 204 206 201 201 200 0.5 1 0 0.5 5 FWHM (mm) 20 100 T 0 -5 P=0.05 threshold for peaks is +/- 5.1467 x (mm) -50 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Delay shift (secs), stimulus average 0 1 3 4 6 7 Subject id, event experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 13.3482mm 1.5 -2 Contour is: magnitude, stimulus average, T statistic > 5 2 1.5 Sd 1 1 0.5 df 200 200 200 200 200 100 200 200 200 200 200 200 200 200 0 0.5 4 FWHM (mm) 20 100 -50 T 0 -2 -4 P=0.05 threshold for peaks is +/- 3.8943 x (mm) 2 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Delay shift (secs), stimulus average 0 1 3 4 6 7 Subject id, block experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 13.5901mm 1.5 -2 Contour is: magnitude, stimulus average, T statistic > 5 2 1.5 Sd 1 1 0.5 df 209 209 214 210 211 210 210 207 212 214 214 210 210 216 0 0.5 4 FWHM (mm) 20 100 -50 T 0 -2 -4 P=0.05 threshold for peaks is +/- 3.983 x (mm) 2 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Delay shift (secs), diff - same speaker 0 1 3 4 6 7 Subject id, event experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 16.9641mm 1.5 -2 Contour is: magnitude, stimulus average, T statistic > 5 2 1.5 Sd 1 1 0.5 df 273 271 276 281 274 136 265 293 264 268 265 264 296 270 0 0.5 4 FWHM (mm) 20 100 -50 T 0 -2 -4 Peaks not significant at P=0.05 x (mm) 2 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Delay shift (secs), diff - same speaker 0 1 3 4 6 7 Subject id, block experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 14.3951mm 1.5 -2 Contour is: magnitude, stimulus average, T statistic > 5 2 1.5 Sd 1 1 0.5 df 201 202 200 206 201 201 200 200 204 204 206 201 205 204 0 0.5 4 FWHM (mm) 20 100 -50 T 0 -2 -4 Peaks not significant at P=0.05 x (mm) 2 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Delay shift (secs), interaction 0 1 3 4 6 7 Subject id, event experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 16.9013mm 1.5 -2 Contour is: magnitude, stimulus average, T statistic > 5 2 1.5 Sd 1 1 0.5 df 278 278 279 280 264 126 277 287 264 272 260 277 264 264 0 0.5 4 FWHM (mm) 20 100 -50 T 0 -2 -4 P=0.05 threshold for peaks is +/- 3.8306 x (mm) 2 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 Delay shift (secs), interaction 0 1 3 4 6 7 Subject id, block experiment 8 9 10 11 12 13 14 15 Mixed effects 2 1 Ef 0 -1 Random /fixed effects sd smoothed 14.4178mm 1.5 -2 Contour is: magnitude, stimulus average, T statistic > 5 2 1.5 Sd 1 1 0.5 df 204 200 207 200 204 205 202 203 202 204 206 201 201 200 0 0.5 4 FWHM (mm) 20 100 -50 T 0 -2 -4 Peaks not significant at P=0.05 x (mm) 2 15 0 10 50 5 -60 -40 -20 0 y (mm) 0 STAT_SUMMARY example: single run, hot-warm Detected by BON and DLM but not by RFT Detected by DLM, but not by BON or RFT