Monday, July 6, 2015

Notes from the OHBM 2015 "Statistical Assessment of MVPA Results" Morning Workshop

Thanks to everyone that attended and gave feedback on the OHBM morning workshop "Statistical Assessment of MVPA Results" that Yaroslav Halchenko and I organized! We've received several requests for slides and materials related to the workshop, so I'll collect them here. It appears that material from the meeting will also be searchable from links on the main OHBM 2015 page. As always, all rights are reserved, and we expect to be fully cited, acknowledged, and consulted for any uses of this material.

I started the workshop off with a tutorial on permutation testing aimed at introducing issues particularly relevant for MVPA (and neuroimaging datasets in general). I'll eventually post a version of the slides, but some of the material is already available in more detail in two PRNI conference papers:
  • Etzel, J.A. 2015. MVPA Permutation Schemes: Permutation Testing for the Group Level. 5th International Workshop on Pattern Recognition in NeuroImaging (PRNI 2015). Stanford, CA, USA. In press, full text here, and in ResearchGate.
  • Etzel, J.A., Braver, T.S., 2013. MVPA Permutation Schemes: Permutation Testing in the Land of Cross-Validation. 3rd International Workshop on Pattern Recognition in NeuroImaging (PRNI 2013). IEEE, Philadelphia, PA, USA. DOI:10.1109/PRNI.2013.44. Full text here, and in ResearchGate.
Next, Johannes Stelzer gave a talk entitled "Nonparametric methods for correcting the multiple comparisons problem in classification-based fMRI", the slides for which are available here.

Then, Nikolaus Kriegeskorte gave a talk entitled "Inference on computational models from predictions of representational geometries", the slides for which are available here.

Finally, Yaroslav Halchenko finished the session with a talk giving an "Overview of statistical evaluation techniques adopted by publicly available MVPA toolboxes", the slides for which are available here

Monday, May 18, 2015

resampling images with wb_command -volume-affine-resample

I often need to resample images without performing other calculations, for example, making a 3x3x3 mm voxel version of an anatomical image with 1x1x1 mm voxels for use as an underlay. This can be done with ImCalc in SPM, but that's a bit annoying, as it requires firing up SPM, and only outputs two-part NIfTI images (minor annoyances, but still).

The wb_command -volume-affine-resample program gets the resampling done at the command prompt with a single long command:

 wb_command -volume-affine-resample d:/temp/inImage.nii.gz d:/temp/affine.txt d:/temp/matchImage.nii CUBIC d:/temp/outImage.nii  

If the wb_command program isn't on the path, run this at the command prompt, from wherever wb_command.exe (or the equivalent for your platform) is installed. A lot of  things need to be specified:
  • inImage.nii.gz is the image you want to resample (for example, the 1x1x1 mm anatomical image)
  • affine.txt is a text file with the transformation to apply (see below)
  • matchImage.nii is the image with the dimensions you want the output image to have - what inImage should be transformed to match (for example, the 3x3x3 mm functional image)
  • CUBIC is how to do the resampling; other options are TRILINEAR and ENCLOSING_VOXEL
  • outImage.nii is the new image that will be written: inImage resampled to match matchImage; specifying a outImage.nii.gz will cause a gzipped NIfTI to be written.
The program writes outImage as a one-file (not a header-image pair) NIfTI. It takes input images as both compressed (i.e., .nii.gz) and uncompressed (i.e., .nii) one-file NIfTIs, but didn't like a header-image pair for input.

You need to specify an affine transform, but I don't want to warp anything so the matrix is all 1s and 0s; just put this matrix into a plain text file (I called it affine.txt):
 1 0 0 0  
 0 1 0 0  
 0 0 1 0  

UPDATE 20 May 2015: Changed the resampling method to CUBIC and added a note that the program can output compressed images, as suggested by Tim Coalson.

Friday, May 15, 2015

MVPA on the surface: to interpolate or not to interpolate?

A few weeks ago I posted about a set of ROI-based MVPA results using HCP images, comparing the results of doing the analysis with the surface or volume version of the dataset. As mentioned there, there hasn't been a huge amount of MVPA with surface data, but there has been some, particularly using the algorithms in Surfing (they're also in pyMVPA and CoSMoMVPA), described by Nikolaas Oosterhof (et al., 2011).

The general strategy in all MVPA (volume or surface) is usually to minimize changing the fMRI timeseries as much as possible; motion correction is pretty much always unavoidable, but is sometimes the only whole-brain image manipulation applied: voxels are kept in the acquired resolution, not smoothed, not slice-time corrected, not spatially normalized to an atlas (i.e., each individual analyzed in their own space, allowing the people to have differently-shaped brains). The hope is that this minimal preprocessing will maximize spatial resolution: since we want to detect voxel-level patterns, let's change the voxels as little as possible.

The surface searchlighting procedure in Surfing follows this minimum-voxel-manipulation strategy, using a combination of surface and volume representations: voxel timecourses are used, but adjacency determined from the surface representation. Rephrased, even though the searchlights are drawn following the surface (using a high-resolution surface representation), the functional data is not interpolated, but rather kept as voxels: each surface vertex is spatially mapped to a voxel, allowing multiple vertices to fall within a single voxel in highly folded areas. Figure 2 from the Surfing documentation  shows this dual surface-and-volume way of working with the data, and describes the voxel selection procedure in more detail. In the way I've described my own searchlight code, the Surfing procedure results in a lookup table (which voxels constitute the searchlight for each voxel) where the searchlights are shaped to follow the surface in a particular way.

It should be possible to do this (Surfing-style, surface searchlights with voxel timecourses) with the released HCP data. The HCP volumetric task-fMRI images are spatially normalized to the MNI atlas, which will simplify things, since the same lookup table can be used with all people, though possibly at the cost of some spatial normalization-caused distortions. [EDIT 17 May 2015: Nick Oosterhof pointed out that even with MNI-normalized volumetric fMRI data, the subject-space surfaces could be used to map adjacent vertices, in which case each person would need their own lookup table. With this mapping, the same i,j,k-coordinate voxel could have different searchlights in different people.]

The HCP task fMRI data is also available as (CIFTI-format) surfaces, which were generated by resampling the (spatially-normalized) voxels' timecourses into surface vertices. The timecourses in the HCP surface fMRI data have thus been interpolated several times, including to volumetric MNI space and to the vertices.

Is this extra interpolation beneficial or not? Comparisons are needed, and I'd love to hear about any if you've tried them. The ones I've done so far are with comparatively large parcels, not searchlights, and certainly not the last word.

grey matter musings

fMRI data is always acquired as volumes,  usually (in humans) with voxels something like 2x2x2 to 4x4x4 mm in size. Some people have argued that for maximum power analyses should concentrate on the grey matter, ideally as surface representations. This strikes me as a bit dicey: fMRI data is acquired at the same resolution all over the brain; it isn't more precise where the brain is more folded (areas with more folding have closer-spaced vertices in the surface representation, so multiple vertices can fall within a single voxel).

But how much of a problem is this? How does the typically-acquired fMRI voxel size compare to the size of the grey matter? Trying to separate out fMRI signals from the grey matter is a very different proposition if something like ten voxels typically fit within the ribbon vs. just one.

Fischl and Dale (2000, PNAS, "Measuring the thickness of the human cerebral cortex from magnetic resonance images") answers my basic question of how wide the grey matter typically is in adults: 2.5 mm. This figure (Figure 3) shows the histogram of grey matter thickness that they found in one person's cortex; in that person, "More than 99% of the surface is between 1- and 4.5-mm thick."

So, it's more typical that the grey matter is one fMRI voxel wide than multiple.  A 4x4x4 mm functional voxel will be wider than nearly all grey matter; most voxels within the grey matter will contain some fractional proportion, not just grey matter. Things are better with 2x2x2 mm acquired voxels, but it will still be the case that a voxel falling completely into the grey matter will be fairly unusual, and even these totally-grey voxels will surrounded on several sides by non-grey matter voxels. To make it concrete, here's a sketch of common fMRI voxel sizes on a perfectly straight grey matter ribbon.

This nearness of all-grey, some-grey, and no-grey voxels is problematic for analysis. An obvious issue is blurring from motion: head motion of a mm or two within a run is almost impossible to avoid, and will totally change the proportion of grey matter within a given voxel. Even if there was no motion at all, the different proportions of grey matter causes problems ("partial volume effects"; see for example): if all the signal came from the grey matter, the furthest-right 2 mm voxels in the image above would be less informative than the adjacent 2 mm voxel which is centered in the grey, just because of the differing proportion grey. Field inhomogeneity effects, scanner drift, slice-time correction, resampling, smoothing, spatial normalization, etc. cause further blurring.

But the cortex grey matter is of course not perfectly flat like in the sketch: it's twisted and folded in three dimensions, like shown here in Figure 1 from Fischl and Dale (2000). This folding leads complicates things further: individual voxels still have varying amounts of grey matter, but can also encompass structures far apart if measured along the surface.

This figure is panels C (left) and D (right) from Figure 2 of Kang et al. (2007, Magnetic Resonance Imaging. Improving the resolution of functional brain imaging), and illustrates some of the "complications". The yellow outline at left is the grey-white boundary on an anatomical image (1x1x1 mm), with two functional voxels superimposed, one in red and one in green (the squares mark the voxels' corners; they had 1.88x1.88x5 mm functional voxels). The right pane shows the same two voxels' locations in a surface flat map (dark areas grey matter, light areas white). In their words, "Although the centers of the filled squares in the corners of the red and green functional voxels in (C) are the same distance apart in the 3-D space and points in the same voxel must be within 5.35 mm, functional activations in the red voxel spread to areas over 30 mm apart on the flat map, while activations in the green voxel remain close to each other."

Volume-to-surface mapping algorithms and processing pipelines attempt to minimize these problems, but there's no perfect solution: acquired voxels will necessarily not perfectly fall within the grey matter ribbon. We shouldn't allow the perfect to be the enemy of the good (no fMRI research would ever occur!) and give up on grey matter-localized analyses entirely, but we also shouldn't discount or minimize the additional difficulties and assumptions in surface-based fMRI analysis.

Tuesday, May 12, 2015

upcoming travels: PRNI and HBM

I'll be traveling a lot next month: attending PRNI June 10-12, then HBM June 14-18. I'll be talking about statistical testing for MVPA at both conferences, focusing on permutation testing for group analyses at PRNI, and a bit more general at our HBM workshop. I hope to meet some of you at one or both of these conferences!

Wednesday, April 22, 2015

ROI-based MVPA: surface and volume comparisons, using HCP data

How should we do a "standard" ROI-based MVPA: on the surface or in the volume? There are a few mass-univariate (e.g., cited in the introduction of Glasser et al., 2013) and searchlight-based (most from comparisons in the literature, but not many, and not for larger ROIs (send some pointers if I just missed 'em).

This post describes a direct comparison of the carrying out the same ROI-based task-classification MVPA using both surface and volume data. It uses the working memory task fMRI data collected in the HCP, which should be some of the best-preprocessed data available now, with the Gordon, et al. (2014) communities as ROIs. Since the Gordon analyses were also done on the surface, I expect these masks to be especially suited for surface analyses. The Gordon parcellation is available perfectly aligned to the HCP volume and surface data, so no transformations (resampling, warping, etc.) are needed.

My intent was to design these analyses to be as "apples-to-apples" as possible, allowing direct comparisons between the surface and volume results. The analyses in this post were carried out in two (non-intersecting) sets of unrelated people (190 in group 1, 160 in group 2) from the 500-subjects HCP data release. I used ten-fold cross-validation in each group (leave-19-out for group 1; leave-16-out for group 2), determining the cross-validation folds ahead of time, so the same people made up the training and testing sets for both the volume and surface analyses and all four pairwise classifications (my intent being to eliminate as many non-surface or volume-related sources of variation as possible). All classifications were with linear SVM, c=1, on the HCP-provided cope images ("parameter estimate images", in my usual parlance), one per person per class (here, 0BK_FACE, OBK_PLACE, 2BK_FACE, 2BK_PLACE).

The analyses consisted of four pairwise classifications in each group of subjects and type of image (surface or volume), taken from the working memory task. Briefly, the working memory task was a blocked version of the n-back, with blocks of either 0-back or 2-back, performed with different categories of pictures. I can thus classify both the picture type used in the block (here, face or place) or the n-back level (0 or 2), and make sensible predictions (since these are well-understood tasks), such that the Visual community should classify face vs. place much better than 0-back vs. 2-back.

These busy graphs show the results. The first graph has the results using the Group 1 subjects, and the second, the Group 2 subjects (i.e., the replications: same analyses in the two groups of people). The accuracies from analyzing the volume data are plotted as solid lines with dots, surfaces, dashed lines with xs. The abbreviations along the x-axis are the 13 communities in the Gordon2014 parcellation. There are two points for each community, left hemisphere on the left side of the line, right hemisphere on the right. The shaded area below 0.6 accuracy is since these accuracies are unlikely to be meaningful (i.e., interpreted as chance).

The classification results are sensible: the Visual community classifies face vs. place extremely well; SMmouth doesn't classify anything; FrontoParietal classifies 0-back vs. 2-back. There's some variation in the details between the two replications (e.g., right-hemisphere None classifies better than left in Group 1, but about the same in Group 2), but the basic pattern of which communities do which classifications is the same in both.

For the surface and volume comparisons, my main impression of the results is of similarity: the accuracies produced from the surfaces and volumes tend to be very close for each community and classification (in the graphs, the dashed and solid lines of each color follow each other). It looks like the difference between the surface and volume versions is less than the difference between whether the analysis was run on Group 1 or 2 (the replications), making me conclude that the surface and volume versions classified  about the same - basically equivalent results either way.

Should this be surprising? Perhaps not: I used the same communities in each analysis, so it's reassuring that they reported basically the same information in the surface and volume versions. The volumetric Gordon community masks closely follow the grey matter (as do the surface reconstructions, of course), so we'd hope they capture the same brain areas. The results would presumably vary more if "lumpier" volumetric ROIs were projected onto the surface.

The number of voxels (volume) in each community is larger than the number of vertices (surface), particularly for larger communities, as shown here (grey line is x=y). This has implications for classification accuracy when using linear SVMs (like I did here), since they can be more likely to detect information when there are more weakly-informative features: more voxels could give an advantage to the volume-based analyses, if they were (even weakly) informative.

We might guess that the surface version would do better than the volume, if the volume included uninformative voxels that weren't assigned a surface vertex, but that doesn't seem to have happened here (perhaps because of the tight grey matter alignment in the volume masks), at least not enough to reduce the accuracy. We might also guess that the surface version would be worse than the volume, for example if preprocessing caused some activity to appear to be in adjacent non-grey-matter areas (which then wouldn't be included in the surface version). {The volumetric BOLD signal is blurred in space during preprocessing (e.g., motion correction and spatial normalization), since preprocessing is usually (including for the HCP) done volumetrically, before the conversion to surfaces.} But in this case, the surface and volume versions came out about the same.

So, can we answer the question I posed at the beginning of this post? How should we do a "standard" ROI-based MVPA: on the surface or in the volume? One set of comparisons can't answer the question definitively, of course, and the HCP data is unusual in many respects (e.g. multiband acquisition, small voxels, specialized preprocessing pipelines). There wasn't an advantage to doing the analyses on the surface here, or much of a disadvantage, since the HCP provides surface versions of the copes. But I'd be hesitant to recommend doing MVPA with fairly large ROIs (like the Gordon communities) on the surface in a new study, particularly if you had to generate the surface files yourself: that's a lot of extra work (making the surface versions) for what might be very little benefit.

Many more comparisons are needed to provide general guidance for when analyses should be done on the surface or volume. I'm particularly interested in trying more comparisons with dilated ROIs: does accuracy improve if voxels in adjacent non-grey matter are included in the mask? If so, we may be better with analyzing the volume. If you run or run across more comparisons, please let me know, and I'll add them here.

The MVPA results here were done in R, similarly to the ROI-based demo. I followed the steps here to read the functional data out of the surface (cifti) files, using Guillaume Flandin's MATLAB library to read the extracted gifti files and get the values for the vertices corresponding to each Gordon parcel. The post is hopefully detailed enough to allow running similar comparisons; I can provide details to replicate exactly if needed.

Monday, April 13, 2015

format conversion: 4dfp to NIfTI, plus setting handedness and headers

This post is a tutorial explaining how to convert fMRI datasets from 4dfp to NIfTI format. Most people not at Washington University in St Louis probably won't encounter a 4dfp dataset, since the format isn't in wide use. But much of the post isn't 4dfp-specific, and goes over ways to check (and correct) the orientation and handedness of voxel arrays. The general steps and alignment tests here should apply, regardless of which image format is being converted to NIfTI.

Converting images between formats (and software programs) is relatively easy ... ensuring that the orientation is set properly in the converted images is sometimes not, and often irritating. But it is very, very worth your time to make sure that the initial image conversion is done properly; sorting out orientation at the end of an analysis is much harder than doing it right from the start.

To understand the issues, remember that NIfTI (and 4dfp, and most other image formats) have two types of information: the 3d or 4d array of voxel values, and how to interpret those values (such as how to align the voxel array to anatomical space, the size of the voxels in mm, etc.), which is stored in a header. This post gives has more background information, as does this one. The NIfTI file specification allows several different ways of specifying alignment, and both left and right-handed voxel arrays. Flexibility is good, I guess, but can lead to great confusion, because different programs have different strategies for interpreting the header fields. Thus, the exact same NIfTI image can look different (e.g. left/right flipped, shifted off center) when opened in different programs. That's what we want to avoid: we want to create NIfTI images that will be read with proper alignment into most programs.

Step 1:  Read the image into R as a plain voxel array

I use the R4dfp R package to read 4dfp images into R, and the oro.nifti package to (read or) write NIfTI images out of R. The R4dfp package won't run in Windows, so needs to be run on a linux box, mac, or within NeuroDebian (oro.nifti runs on all platforms).

 in.4dfp <- R4dfp.Load(paste0(in.path,;  
 vol <- in.4dfp[,,,];  

The 4dfp image fname at location in.path is read into R with the first command, and the voxel matrix extracted with the second. You can check properties of the 4dfp image by checking its fields, such as in.4dfp$dims and in.4dfp$center. The dimension of the extracted voxel array should match the input 4dfp fields (in.4dfp$dims == dim(vol)).

Step 2:  Determine the handedness of the input image and its alignment

Now, look at the extracted voxel array: how is it orientated? My strategy is to open the original image in the program whose display I consider to be the ground truth, then ensure the matrix in R matches that orientation. For 4dfp images, the "ground truth" program is fidl; often it's the program which created the input image. I then look at the original image and find a slice with a clear asymmetry, and display that slice in R with image.

Here's slice k=23 (green arrow) of the first timepoint in the image (blue arrow). The greyscale image is the view in fidl which we consider true: the left anterior structure (white arrow) is "pointy". The R code image(vol[,,23,1])) displays this same slice, using hot colors (bottom right). We can see that it's the correct slice (I truncated the side a bit in the screen shot), but the "pointy" section is at the bottom right of the image instead of the top left. So, the voxel array needs flipped left/right and anterior/posterior.

Step 3:  Convert and flip the array as needed

This doFlip R function reorders the voxel array. It takes and returns a 3d array, so should be run on each timepoint individually. This takes a minute or so to run, but I haven't bothered speeding it up, since it only needs to be run once per input image. If you have a faster version, please share!

 # flip the data matrix in the specified axis direction(s). Be very careful!!!  
 doFlip <- function(inArray, antPost=FALSE, leftRight=FALSE) {  
   if (antPost == FALSE & leftRight == FALSE) { stop("no flips specified"); }  
   if (length(dim(inArray)) != 3) { stop(print("not three dimensions in the array")); }  
   outvol <- array(NA, dim(inArray));  
   IMAX <- dim(inArray)[1];  
   JMAX <- dim(inArray)[2];  
   KMAX <- dim(inArray)[3];  
   for (k in 1:KMAX) {  
     for (j in 1:JMAX) { # i <- 1; j <- 1; k <- 1;  
       for (i in 1:IMAX) {  
         if (antPost == TRUE & leftRight == FALSE) { outvol[i,j,k] <- inArray[i,(JMAX-j+1), k]; }  
         if (antPost == TRUE & leftRight == TRUE) { outvol[i,j,k] <- inArray[(IMAX-i+1),(JMAX-j+1), k]; }  
         if (antPost == FALSE & leftRight == TRUE) { outvol[i,j,k] <- inArray[(IMAX-i+1), j, k]; }  
  # put the array to right-handed coordinate system, timepoint-by-timepoint.  
  outvox <- array(NA, dim(vol));  
  for (sl in 1:dim(vol)[4]) { outvox[,,,sl] <- doFlip(vol[,,,sl], TRUE, TRUE); }  

The last three code call the function  (TRUE, TRUE specifies flipping both left-right and anterior-posterior), putting the flipped timepoint images into the new array outvox. Now, when we look at the same slice and timepoint with the image command (image(outvox[,,23,1]))), it is orientated correctly.

Step 4:  Write out as a NIfTI image, confirming header information

A 3 or 4d array can be written out as a NIfTI image without specifying the header information (e.g. with writeNIfTI(nifti(outvox), fname), which uses the oro.nifti defaults). Such a "minimal header" image can be read in and out of R just fine, but will almost certainly not be displayed properly when used as an overlay image in standard programs (e.g., on a template anatomy in MRIcroN) - it will probably be out of alignment. The values in the NIfTI header fields are interpreted by programs like MRIcroN and the Workbench to ensure correct registration.

Some of the NIfTI header fields are still mysterious to me, so I usually work out the correct values for a particular dataset iteratively: writing a NIfTI with my best guess for the correct values, then checking the registration, then adjusting header fields if needed.

  out.nii <- nifti(outvox, datatype=64, dim=dim(outvox), srow_x=c(3,0,0,in.4dfp$center[1]),   
      srow_y=c(0,3,0,in.4dfp$center[2]), srow_z=c(0,0,3,in.4dfp$center[3]))   
  out.nii@sform_code <- 1    
  out.nii@xyzt_units <- 2; # for voxels in mm    
  pixdim(out.nii)[1:8] <- c(1, 3,3,3, dim(outvox)[4], 1,1,1);   
   # 1st slot is qfactor, then mm, then size of 4th dimension.   
  writeNIfTI(out.nii, paste0(out.path,    

These lines of code write out a 4d NIfTI with header values for my example image. The input image has 3x3x3 mm voxels, which is directly entered in the code, and the origin is read out of the input fields (you can also type the numbers in directly, such as srow_x=c(3,0,0,72.3)). The image below shows how to confirm that the output NIfTI image is orientated properly: the value of the voxel under the crosshairs is the same (blue arrows) in the original 4dfp image in fidl, the NIfTI in R, and the NIfTI in MRIcroN. The red arrows point out that the voxel's i,j,k coordinates are the same in MRIcroN and R, and the left-right flipping matches (yellow arrow).

Here's another example of what a correctly-aligned file looks like: the NIfTI output image is displayed here as an overlay on a template anatomy in MRIcroN (overlay is red) and the Workbench (overlay is yellow). Note that the voxel value is correct (blue), and that the voxel's x,y,z coordinates (purple) match between the two programs. If you look closely, the voxel's i,j,k coordinates (blue) are 19,4,23 in MRIcroN (like they were in R), but 18,3,22 in the Workbench. This is ok: Workbench starts counting from 0, R and MRIcroN from 1.

concluding remarks

It's straightforward to combine the R code snippets above into loops that convert an entire dataset. My strategy is usually to step through the first image carefully - like in this post - then convert the images for the rest of the runs and participants in a loop (i.e., everyone with the same scanning parameters). I'll then open several other images, confirming they are aligned properly.

This last image is a concrete example of what you don't want to see: the functional data (colored overlay) is offset from the anatomical template (greyscale). Don't panic if you see this: dive back into the headers.