Group Analysis with AFNI Programs
Introduction
Most of the material and notations are from Doug WardÕs manuals for the programs 3dttest, 3dANOVA, 3dANOVA2, 3dANOVA3, and 3dRegAna, and from Gang ChenÕs manual for 3dANOVA4
Documentation available with the AFNI distribution
Lots of stuff (theory, examples) therein
Software and documentation files are based on these books:
Applied Linear Statistical Models by Neter, Wasserman, and Kutner (4th edition)
Applied Regression Analysis by Draper and Smith (3rd edition)
General steps
Smoothing (3dmerge -1blur_fwhm)
Normalization (3dcalc)
Deconvolution/Regression (3dDeconvolve)
Co-registration of individual analyses to common ÒspaceÓ (adwarp -dxyz)
Group analysis (3dttest, 3dANOVA, É)
Post-analysis (AlphaSim, conjunction analyses, É)
Interpretation and  Thinking

"Data Preparation:"
Data Preparation: Spatial Smoothing
Spatial variability of both FMRI activation and the Talairach transform (the common space) can result in little or no overlap of function between subjects.
Data smoothing is used to reduce this problem.
Leads to loss of spatial resolution, but that is a price to be paid with the Talairach transform (or any current technique that does inter-subject anatomical alignments)
In principle, smoothing should be done on time series data, before data fitting (i.e., before 3dDeconvolve or 3dNLfim, etc.)
Otherwise one has to decide on how to smooth statistical parameters.
In statistical data sets, each voxel has a multitude of different parameters associated with it like a regression coefficient, t-statistic, F-statistic, etc.
Combining some statistical parameters across voxels would result in parameters with unknown distributions
It is OK to blur percent signal change values that come out of the regression analysis, since these numbers depend linearly on the input data (unlike the F- and t-statistics)
Blurring in 3D is done using 3dmerge with the -1blur_fwhm option
Blurring on the surface is done with program SurfSmooth

"Data Preparation:"
Data Preparation: Parameter Normalization
Parameters quantifying activation must be normalized before group comparisons.
FMRI signal amplitude varies for different subjects, runs, scanning sessions, regressors, image reconstruction software, modeling strategies, etc.
Amplitude measures (regression coefficients) can be turned to percent signal change from baseline (do it before the individual analysis in 3dDeconvolve).
Equations to use with 3dcalc to calculate percent signal change
100 bi / b0 (basic formula)
100 bi / b0 * c  (mask out the outside of the brain)
bi  = coefficient for regressor i (output from 3dDeconvolve)
b0 = baseline estimate (output from 3dTstat -mean)
c  = threshold value generated from running 3dAutomask -dilate
This will be included into 3dDeconvolve in a future release
Other normalization methods, such as z-score transformations of statistics, can also be used.

"Data Preparation:"
Data Preparation: Co-Registration (AKA ÒSpatial NormalizationÓ)
Group analyses are performed on a voxel-by-voxel basis
All data sets used in the analysis must be aligned and defined over the same spatial domain.
Talairach domain for volumetric data
Landmarks for the transform are set on high-res. anatomical data using AFNI
Functional data volumes are then transformed using AFNI interactively or adwarp from command line (use option -dxyz with about the same resolution as EPI data Ñ do not use the default 1 mm resolution!)
Standard meshes and spherical coordinate system for surface data
Surface models of the cortical surface are warped to match a template surface using Caret/SureFit (http://brainmap.wustl.edu) or FreeSurfer (http://surfer.nmr.mgh.harvard.edu)
Standard-mesh surface models are then created with SUMA (http://afni.nimh.nih.gov/ssc/ziad/SUMA) to allow for node-based group analysis using AFNIÕs programs
Once data is aligned, analysis is carried out voxel-by-voxel or node-by-node
The percent signal change from each subject in each task/stimulus state are usually the numbers that will be compared and contrasted
Resulting statistics (voxel-wise or node-wise) can then be displayed in AFNI and/or SUMA

"Overview of Statistical Testing of..."
Overview of Statistical Testing of Group Datasets with AFNI programs
Parametric Tests:
Assume data are normally distributed (Gaussian)
3dttest  (paired, unpaired)
3dANOVA (or 3dANOVA2 or 3dANOVA3)
3dRegAna (regression, unbalanced ANOVA, ANCOVA)
3dANOVA4 = Matlab script for one-, two-, three- and four-way ANOVA
Non-parametric analyses:
No assumption of normality
Tends to be less sensitive to outliers (more robust)
3dWilcoxon (~t-test paired)
3dMannWhitney (~t-test unpaired)
3dKruskalWallis (~3dANOVA)
3dFriedman (~3dANOVA2)
Permutation test
Less sensitive and less flexible than parametric tests
In practice, seems to make little difference
Probably because number of datasets and subjects is usually small (hard to tell if data is non-Gaussian when only have a few sample points)

"t-Tests"
t-Tests  [starting easy, but contains most of the ideas]
Program 3dttest
Used to test if the mean of a set of values is significantly different from a constant (usually 0) or the mean of another set of values.
Assumptions
Values in each set are normally distributed
Equal variance in both sets
Values in each set are independent Þ unpaired t-test
Values in each set are dependent Þ paired t-test
Example: 20 subjects are tested for the effects of 2 drugs A and B
Case 1: 10 subjects were given drug A and the other 10 subjects given drug B.
Unpaired t-test is used to test: mA = mB? (mean response is different?)
Equivalent to one-way ANOVA with between-subjects design of equal sample size Þ can also run 3dANOVA (treating subjects as multiple measurements)
Case 2: 20 subjects were given both drugs at different times.
Paired t-test is used to test: mA = mB?
Case 3: 20 subjects were given drug A.
t-test is used to test if drug effect is significant at group level: mA = 0?

Unpaired 2 Sample t-Test: Cartoon Data
Paired t-Test: Cartoon Data
"1-Way ANOVA"
1-Way ANOVA
Program 3dANOVA
Determine whether treatments (levels) of a single factor (independent parameter) has an effect on the measured response (dependent parameter, like FMRI percent signal change due to some stimulus).
Examples of factor: subject type, task type, task difficulty, drug type, drug dosage, etc.
Within a factor are levels: different sub-categorizations
Example: factor=subject type; level 1=normals, level 2=patients with mild symptoms, level 3=patients with severe symptoms
The various AFNI ANOVA programs differ in the number of factors they allow: 3dANOVA allows 1 factor,  comprising up to 100 levels
Assumptions
Values are normally distributed
No assumptions about relationship between dependent and independent variables (e.g., not necessarily linear)
Independent variables are qualitative
Can also use 3dttest if there are only two levels
The 1-way 3dANOVA analysis is a generalization to multiple levels of an unpaired 3dttest (for generalization of paired, wait for 3dANOVA2)
Example: r different types of subjects performed the same task in the scanner

"Null Hypothesis:"
  Null Hypothesis:       H0 :  m1 = m2 = É = mr
i.e., subject type has no effect on mean signal in this voxel
Alternative Hypothesis: Ha : not all mi are equal
i.e., at least one subject type had a different mean FMRI signal
3dANOVA is effectively a generalization of the unpaired t-test to multiple columns of data (a further refinement will be introduced with 3dANOVA3)
As such, 3dANOVA is probably not appropriate when comparing results of different tasks on the same subjects (need a generalization of the paired t-test: 3dANOVA2)
Difference from doing unpaired t-tests on pairs of columns: variance estimates are all pooled together, increasing the denominatorial degrees of freedom
Assumption is that data fluctuations in each column have same variance

"ANOVA:"
ANOVA: Which levels had an effect or were different from one another?
Usually, just knowing that there is a main effect (some of the means are different, but no information about which ones) isnÕt enough, so there is a number of options to let you look for more detail
Which treatment means (mi ) are ­ 0 ?
e.g., is the response of subjects in level #3 different from 0 ?
t-statistic with option -mean in 3dANOVA
Similar to using 3dttest -base1 0  (single sample test) to test only the data from those subjects
Which treatment means are different from each other ?
e.g., is the response of subjects in level #3 different from those in level #2 ?
t-statistic with option -diff in 3dANOVA
Similar to using 3dttest (unpaired) between the data from these sets of subjects
Which linear combination of means (contrasts) are ­ 0 ?
e.g., is the average response of subjects in level #1 different from the combined average of subjects in levels #2 and #3 ?
t-statistic with option -contr in 3dANOVA

"2-Way ANOVA:"
2-Way ANOVA: test for effects of two independent factors on measurements
This is a fully crossed analysis: all combinations of factor levels are measured
In particular, if one factor is ÒsubjectÓ, then all subjects are tested in all levels of the other factor
Program is limited to balanced designs: Must have same number of measurements in each ÒcellÓ (combinations of factor levels)
Example: Stimulus type for factor A and subject for factor B
Each subject is a level within factor B (1 measurement per cell)
This is a fixed effect « random effect model = Òmixed effectÓ model
Example: Stimulus type for factor A and drug treatment for factor B
Each subject is an independent measurement for both factors, all levels
This is a fixed effect « fixed effect model
If you also want to treat subject as a separate factor, need 3dANOVA3
Example: Stimulus type for factor A, stimulus day for factor B
With one fixed subject, for a longitudinal study (e.g., training between scan days)
This also is a fixed effect « fixed effect model
Again, multiple subjects could be treated as independent measurements in 3dANOVA2, or as a third factor in 3dANOVA3

"Choose between two types of..."
Choose between two types of analysis for each factor: fixed and random effects
Fixed effects factor = differences between levels in this factor are modeled as deterministic differences in the mean measurements (as in 3dANOVA and 3dttest)
Useful for most categories under the experimenterÕs control or observation
Allows same type of statistics as 3dANOVA:
factor main effect (are all the mean activations of each level in this factor the same?)
differences between level pairs (e.g., level #2 same as #3?)
more complex contrasts (e.g., average of levels #1 and #2 same as level #3?)
If both factors are modeled as fixed effects with multiple measurements (e.g., subjects):
Can also test for interaction between the factors
Are there any combinations of factor levels whose means Òstick outÓ [e.g., mean of cell #(A1,B2) differs from (#A1 mean)+(#B2 mean)]?
Example: A=stimulus type, B=drug type; then cell #(A1,B2) is FMRI response (in each voxel) to stimulus #1 and drug #2
Interaction test would determine if any individual combination of drug type and stimulus type was abnormal
e.g., if stimulus #1 averages a high response, and drug #2 averages no effect on response, but when together, value in cell #(A1,B2) averages small
i.e., Effect of one factor (stimulus) depends on level of other factor (drug)
no interaction means the effects of the factors are always just additive
Inter-factor contrasts can then be used to test individual combinations of cells to determine which cell(s) the interaction comes from

"Random effects factor = differences..."
Random effects factor = differences between levels in this factor are modeled as random fluctuations
Useful for categories not under experimenterÕs control or observation
In FMRI, is especially useful for subjects; a good rule is
treat subjects as a separate random effects factor rather than
as multiple independent measurements inside fixed-effect factors
In such a case, usually have 1 measurement per cell (each cell is the combination of a level from the other factor with 1 subject)
This is sometimes called a Òrepeated measures ANOVAÓ, when we have multiple measurements on each subject (in this case, across different stimulus classes)
Treating subjects as a random factor in a fully crossed analysis is a generalization of the paired t-test
intra-subject and inter-subject data variations are modeled separately
which can let you detect small intra-subject changes due to the fixed-effect factors that might otherwise be overwhelmed by larger inter-subject fluctuations
Main effect for a random effects factor tests if fluctuations among levels in this factor have additional variance above that from the other random fluctuations in the data
e.g., Are inter-subject fluctuations bigger than intra-subject fluctuations?
Not usually very interesting when random factor = subject
It is hard to think of a good FMRI example where both factors would be random
3dANOVA2: Usually have 1 fixed factor and 1 random factor = mixed effects analysis

"NOTE WELL:"
NOTE WELL: Must have same number of observations (Òn Ó) in each cell
 Can use 3dRegAna if you donÕt have the same number of values in each cell (program usage is much more complicated)

"3dANOVA2:"
3dANOVA2: A test case
Michael S. Beauchamp, Kathryn E. Lee, James V. Haxby, and Alex Martin, fMRI Responses to Video and Point-Light Displays of Moving Humans and Manipulable Objects, Journal of Cognitive Neuroscience, 15: 991-1001  (2003).
Purpose is to study the organization of brain responses to different types of complex visual motion (the 4 levels within factor A) from 9 subjects (the levels within factor B)
Data from 3 of the subjects, and scripts to process it with AFNI programs, are available in AFNI HowTo #5 (hands-on)
Available for download at the AFNI web site: http://afni.nimh.nih.gov/afni/doc/howto/
If you want all the data, it is at the FMRI Data Center at Dartmouth: http://www.fmridc.org
Or at least, it should be (but they havenÕt posted it yet for some reason)

"Stimuli:"
Stimuli: Video clips of the following
  Human whole-body motion (HM)

Slide 18
"Data Processing Outline"
Data Processing Outline
Image registration with 3dvolreg
Images smoothed (4 mm FWHM) with 3dmerge
IRF for each of the 4 stimuli were obtained using 3dDeconvolve
Regressor coefficients (IRFs) were normalized to percent signal change (using 3dcalc)
An average activation measure was obtained by averaging IRF amplitude from the 4th through the 10th second of the response (using 3dTstat)
Capturing the positive blood-oxygenation level dependent response but not any post-stimulus undershoot
These activation measures will be the measurements in the ANOVA table
After each subjectÕs results are warped to Talairach coordinates, using adwarp program
3dANOVA2 was carried out with:
 Factor A, fixed effects:      levels = HM, TM, HP, TP (4 types of stimuli)
 Factor B, random effects: levels = 9 subjects
 1 measurement per cell

"3dANOVA2 script"
3dANOVA2 script
3dANOVA2 -type 3 -alevels 4 -blevels 9 \
-dset 1 1 ED+tlrc'[0]' -dset 2 1 ED+tlrc'[1]' \
-dset 3 1 ED+tlrc'[2]' -dset 4 1 ED+tlrc'[3]' \
-dset 1 2 EE+tlrc'[0]' -dset 2 2 EE+tlrc'[1]' \
-dset 3 2 EE+tlrc'[2]' -dset 4 2 EE+tlrc'[3]' \
É É
-dset 1 9 FN+tlrc'[0]' -dset 2 9 FN+tlrc'[1]' \
-dset 3 9 FN+tlrc'[2]' -dset 4 9 FN+tlrc'[3]' \

-amean 1 TM -amean 2 HM -amean 3 TP  -amean 4 HP \
-acontr 1 1 1 1 AllAct \
-acontr -1 1 -1 1 HvsT \
-acontr 1 1 -1 -1 MvsP \
-acontr 0 1 0 -1 HMvsHP \
-acontr 1 0 -1 0 TMvsTP  \
-acontr 0 0 -1 1 HPvsTP \
-acontr -1 1 0 0  HMvsTM \
-acontr  1 -1 -1 1 Inter \
-fa StimEffect \
  -bucket AvgANOVA

"3dANOVA2:"
3dANOVA2: specifying input datasets
3dANOVA2 -type 3 -alevels 4 -blevels 9 \
-dset 1 1 ED+tlrc'[0]' -dset 2 1 ED+tlrc'[1]' \
-dset 3 1 ED+tlrc'[2]' -dset 4 1 ED+tlrc'[3]' \
-dset 1 2 EE+tlrc'[0]' -dset 2 2 EE+tlrc'[1]' \
-dset 3 2 EE+tlrc'[2]' -dset 4 2 EE+tlrc'[3]' \
É É
-dset 1 9 FN+tlrc'[0]' -dset 2 9 FN+tlrc'[1]' \
-dset 3 9 FN+tlrc'[2]' -dset 4 9 FN+tlrc'[3]' \

"3dANOVA2:"
3dANOVA2: specifying which statistics to output
   3dANOVA2 -type 3 -alevels 4 -blevels 9 ÉÉ \
  -amean 1 TM -amean 2 HM -amean 3 TP -amean 4 HP  \
  -acontr 1 1 1 1 AllAct \
  -acontr -1 1 -1 1 HvsT \
  -acontr 1 1 -1 -1 MvsP \
  -acontr 0 1 0 -1 HMvsHP \
  -acontr 1 0 -1 0 TMvsTP  \
  -acontr 0 0 -1 1 HPvsTP \
  -acontr -1 1 0 0  HMvsTM \
  -acontr  1 -1 -1 1 Inter \
  -fa StimEffect \
  -bucket AvgANOVA
-amean 1 TM:  estimate mean of factor A, level 1 and label it TM in the output dataset
-acontr :  specifies contrast matrix and label in output dataset
1  1  1  1:  all of factor A's levels summed = 0?
-1  1 -1  1:  contrast between human and tools (HM + HP)  Р (TM + TP)
1  1 -1 -1:  contrast between motion and points (HM + TM) Ð (HP + TP)
0  1  0 -1:  contrast between human motion and points (HM Ð HP)
É É
-fa StimEffect:  F-statistic for main effect of factor A (any differences among stimuli?)
-bucket AvgANOVA: prefix of output dataset containing statistical results

"3dANOVA2:"
3dANOVA2: viewing results
Main effect: Regions showing presence of differences in activation due to changes in stimulus type (which differences must be determined via later contrasts)
view StimEffect sub-bricks for function and threshold (F-stat = 15, p =10-5)
Factor Means: Activation in response to each category
view TM, HM, etc. sub-bricks (t-stat = 10.6, p = 10-10)
all categories appear to activate same areas
Choose AllAct sub-bricks for finding regions activated by at least one of the stimuli
this region of activation is often used to select an ROI which is examined for subtler effects
Choose HvsT (human versus tools) sub-bricks
note small range of t-values (subtler effects, if any)
lower t-stat threshold to 4, p ~ 5x10-4
might want to restrict hypothesis testing to region activated by stimuli
Look for interactions that might complicate your fairy tale (AKA hypothesis)
view the Inter sub-bricks to determine if some areas for which the contrast (TM+HP)Ð(HM+TP) is significant
Hopefully youÕll find few/none, or be prepared to explain such activations

"3-Way ANOVA:"
3-Way ANOVA: 3dANOVA3 (again, balanced designs only)
Read the manual first and understand what options are available
It is important to understand 2-way ANOVA before moving up to the big time show!
Has several fixed effects and random effects combinations
Has new concept: nested design (vs. fully crossed design)
Nested design is for use when you have 2 fixed effects factors and 1 random effects factor where the subjects for the random effects factor depend on one of the fixed effect factors; example:
factor A = subject type; level #1=normal, #2=genotype Q, #3=genotype R
factor B = stimulus type; levels #1Ð4=different types of videos
factor C = subject; levels #1Ð10 = 30 different subjects, 10 in each of the factor A levels; C is ÒnestedÓ inside A
Nested design is a mixture of unpaired and paired tests
Will be like ÒpairedÓ for tests across stimulus type (factor B levels)
Will be like ÒunpairedÓ across subject types (factor A levels)
Fully crossed design is when the subjects are common across the other factors
As was said before, un-nested design is a generalization of paired t-test
Treating the subjects correctly is a crucially important decision
Unlike 3dANOVA2, 3dANOVA3 does not currently allow for arbitrary contrasts between random cells in different factors/different levels

"4-Way ANOVA:"
4-Way ANOVA: ready to rock-n-roll (for the daring and intrepid)
Interactive Matlab script
Can run both crossed and nested (i.e., subject nested into gender) design
Heavy duty computation + Matlab: expect to take 10s of minutes to hours
Same script can also do ANOVA, ANOVA2, and ANOVA3 analyses
Includes contrast tests across all factors
At present, must have a balanced design with no missing data
equal number of entries in each cell
can be a problem when studying patients (e.g., hard to find some genotypes)
Working now to implement more options, such as
ANCOVA (ANOVA plus regression with continuous covariates; e.g., age)
Recent news: now working!!
unbalanced designs (uneven numbers of entries in cells, or levels in factors)
Recent news: now working for some cases!
missing data (e.g., some subjects couldnÕt perform certain tasks)
Goal: be a user-friendly alternative to running 3dRegAna for most complicated analyses of group datasets
Goal: once program is stabilized, re-write in C for speed and independence from the commercial product Matlab

Slide 26
Further Directions for Group Analysis Research and Software
In a mixed effects model, ANOVA cannot deal with unequal variances in the random factor between different levels of a fixed factor
Example: 2-way layout, factor A=stimulus type (fixed effect), factor B=subject (random effect)
 As seen earlier, ANOVA can detect differences in means between levels in A (different stimuli)
But if the measurements from different stimuli also have significantly different variances (e.g., more attentional wandering in one task vs. another), then the ANOVA model for the signal is wrong
In general, this ÒheteroscedasticityÓ problem is a difficult one, even in a 2-sample t-test; there is no exact F- or t-statistic to test when the means and the variances might differ simultaneously
Although ANOVA does allow somewhat for intra-subject correlations in measurements, it is not fully general
Example: 2-way layout as above, 3 stimulus types in factor A; general      correlation matrix between the 3 different types of responses is                            but ANOVA only properly deals with the  case r12=r13=r23                                       (recall we are assuming subject effects are random; this is the                                correlation matrix for the intra-subject random responses).
Possible solution: general linear-quadratic minimum variance mixed effects modeling
A statistical theory not yet much applied to FMRI data (but it will be, someday)
Questions of sample size (number of subjects needed) will surely arise

"Conjunction Junction:"
Conjunction Junction: WhatÕs Your Function?
The program 3dcalc is a general purpose program for performing logic and arithmetic calculations
 Command line is of the format
3dcalc -a Dset1 -b Dset2 ... -expr Ò(a * b ...)Ó
 Some expressions can be used to select voxels with values v meeting certain criteria:
Find voxels where v ³ th and mark them with value=1
    expression =   step (v Ð th)   (result is 1 or 0)
In a range of values: thmin ² v ² thmax
    expression =   step (v Ð thmin) * step (thmax - v)
Exact value: v = n
      expression =   equals(v Ð n)
 Create masks to apply to functional datasets
Two values both above threshold (e.g., active in both tasks; ÒconjunctionÓ)
    expression =   step(v-A)*step(w-B)

"Regression Analysis:"
Regression Analysis: 3dRegAna
Simple linear regression:
Y = b0 + b1X1,+ e
where Y represents the FMRI measurement (i.e., percent signal change) and X is the independent variable (i.e., drug dose)
Multiple linear regression:
Y = b0 + b1X1 + b2X2 + b3X3 + É+ e
Regression with qualitative and quantitative variables (ANCOVA)
i.e., drug dose (5mg, 12mg, 23mg, etc.) is quantitative while drug type (Nicotine, THC, Cocaine) or age group (young vs. old) or genotype is qualitative, and usually called dummy (or indicator) variable
ANOVA with unequal sample sizes (with indicator variables)
Polynomial regression:
Y = b0 + b1X1 + b2X12 + É + e
Linear regression: model is a linear function of its unknowns bi  , NOT its independent variables Xi
Not for fitting time series, use 3dDeconvolve (or 3dNLfim) instead

"F-test for Lack of..."
F-test for Lack of Fit (lof)
If multiple measurements are available (and they should be), a Lack Of Fit (lof) test is first carried out.
Hypothesis:
H0: E(Y) = b0 + b1X1 + b2X2 + É,+ bp-1Xp-1
Ha: E(Y) ­ b0 + b1X1 + b2X2 + É,+ bp-1Xp-1
Hypothesis is tested by comparing the variance of the modelÕs lack of fit to the measurement variance at each point (pure error).
If Flof  is significant then model is inadequate. STOP HERE.
Reconsider independent variables, try again.
If Flof  is insignificant then model appears adequate, so far.
It is important to test for the lack of fit:
The remainder of the analysis assumes an adequate model is used
You will not be visually inspecting the goodness of the fit for thousands of voxels!

"Test for Significance of Linear..."
Test for Significance of Linear Regression
This is done by testing whether additional parameters significantly improve the fit
For simple case
Y = b0 + b1X1 + e
H0:  b1 = 0
H1:  b1 ­ 0
For general case
Y = b0 + b1X1 + b2X2 +  É + bq-1Xq-1 +  bqXq + É + bp-1Xp-1 + e
H0: bq = bq+1 = ... = bp-1 = 0
Ha: bk ­ 0, for some k, q ² k ² p-1
Freg is the F-statistic for determining if the Full model significantly improved on the reduced model
NOTE: This F-statistic is assumed to have a central F-distribution. This is not the case when there is a lack of fit

"3dRegAna:"
3dRegAna: Other statistics
How well does model fit data?
R2 (coefficient of multiple determination) is the proportion of the variance in the data accounted for by the model 0 ² R2 ² 1.
i.e., if R2 = 0.26 then 26% of the dataÕs variation about their mean is accounted for by the model. So this might indicate the model, even if significant, might not be that useful (depends on what use you have in mind)
Having said that, you should consider R2 relative to the maximum it can achieve given the pure error which cannot be modeled. [cf. Draper & Smith, chapter 2].
Are individual parameters bk significant?
t-statistic is calculated for each parameter
helps identify parameters that can be discarded to simplify the model
R2 and t-statistic are computed for full (not reduced) model

Examples from Applied Regression Analysis by Draper and Smith (third edition)
"3dRegAna:"
3dRegAna: Qualitative Variables (ANCOVA)
Qualitative variables can also be used
i.e., WeÕre modeling the response amplitude to a stimulus of varying contrast when subjects are either young, middle-aged or old.
X1 represents the stimulus contrast (quantitative): continuous covariate
Create indicator variables X2 and X3 to represent age:
X2 = 1 if subject is middle-aged
= 0 otherwise
X3 = 1 if subject is old (i.e., at least 1 year older than Bob Cox)
= 0 otherwise
Full Model (no interactions between age and contrast)
Y = b0 + b1X1 + b2X2 + b3X3 + e
E(Y) = b0 + b1X1   for young subjects
E(Y) = ( b0 + b2 ) + b1X1 for middle-aged subjects
E(Y) = ( b0 + b3 ) + b1X1 for old subjects
Full Model (with interactions between age and contrast)
Y = b0 + b1X1 + b2X2 + b3X3 + b4X2X1 + b5X3X1 + e
E(Y) = b0 + b1X1   for young subjects
E(Y) = ( b0 + b2 )  + ( b1 + b4 )X1 for middle-aged subjects
E(Y) = ( b0 + b3 ) + ( b1 + b5 )X1 for old subjects
Will be easier to run analysis in Matlab script for 3dANOVA4, when ready!

"3dRegAna:"
3dRegAna: ANOVA with unequal samples
3dANOVA2 and 3dANOVA3 do not allow for unequal samples in each combination of factor levels
Can use 3dRegAna to look for main effects and interactions
The analysis method involves the use of indicator variables so it is practical for small for small number (~3) of factor levels
Details are in the 3dRegAna manual
method is significantly more complicated than running ANOVA; you must understand the math
avoid this, if you can, especially if you have more than 4 factor levels or more than 2 factors
Interactions hard to interpret, and contrast tests unavailable
Will be easier to run analysis in Matlab script for 3dANOVA4, when ready!