DTI Analysis: Soup to Nuts Playlist

Instead of going through each DTI analysis step individually, I've collated everything into a Youtube playlist down below. Just remember that we are using data from the FSL practical course here, and also remember that suing somebody for giving out bad advice, although it is admittedly an easy way to become fantastically wealthy, won't necessarily make you happier.

In any case, just to briefly go over the rest of the steps: After correcting for magnetic field distortions and eddy currents, tensors are fitted using the dtifit command (or simply going through the FDT menu in the FSL interface). Once this has been done for each subject, a series of TBSS tools are used, each one prefixed by "tbss"; for example, tbss_1_preproc, tbss_2_reg, and so on. (You can find all of these in the $FSLDIR/bin directory, and if you have a good handle on Unix programming, you can inspect the code yourself.) After you have run all of those for your dataset, you set up the appropriate experimental design and contrasts, and use the randomise tool to perform statistics in your tractography mask.

Keep in mind that this is just beginner's material; and that if you were to compare your DTI competence to dating, if would be like you were still in that awkward teenager phase, unable to talk to anybody or make eye contact.

However, much of this material has already been covered in other blogs and online resources, provided by several other highly talented scientists and researchers, and - as much as I constantly fantasize about establishing a monopoly over neuroimaging information - there is no practical way to destroy them all.

Therefore, instead of posting redundant information, I highly recommend checking out an ongoing step-by-step series on TORTOISE by Peter Molfese, which you can compare to your results with FSL, and another blog dedicated to diffusion imaging, diffusion-imaging.com. The latter site covers virtually every piece of diffusion imaging software and pipeline out there, and is a good place to start.

DTI Analysis, Steps 1 & 2: Distortion Correction and Eddy Correction

DTI data comes into this world just as we do - raw, unprocessed, unthresholded, and disgusting. Today's parents are much too easy on their children; back in my day, boys and girls were expected to correct their own erratic motion artifacts, to strip their own skulls when too much cranium started sprouting around their cortex, and reach significance through their own diligence and hard work. "No dinner for you until you've gotten rid of those filthy distortion currents on your hands, young man!" was a common saying around my household.

Unfortunately, these days we live in a society increasingly avoiding taking responsibility for their own actions, filled with people more and more unwilling to strip their own skulls. (Even though that would save neuroimaging researchers countless hours of preprocessing.) This trend is the result of several complex cultural and sociological factors, by which I mean: Lawyers. Things have gotten so bad that you could, to take a random example, potentially sue someone posting advice on an unlicensed website if following those instructions somehow messed up your data.

This actually happened to a friend of mine who publishes the nutrition blog tailored for Caucasian males, White Man's Secret. After one of his readers formed kidney stones as a result of following advice to drink a two-liter of Mello Yello every day, the author was sued for - and I quote from the court petition - "Twelve Bazillion Dollars."

In any case, DTI data takes some time to clean up before you can do any statistics on it. However, the preprocessing steps are relatively few, and are analogous to what is done in a typical FMRI preprocessing pipeline. The steps are to identify and correct any distortions during the acquisition, and then to correct for eddy currents.

The first step, distortion correction, can be done with FSL's topup command. Note that this will require two diffusion weighted images, one with the encoding direction in anterior-to-posterior direction (AP) and one encoded in the posterior-to-anterior direction (PA). You will also need a textfile containing information about the encoding directions and the total readout time (roughly the amount of time between the first echo and the last echo); for example, let's say we have a text file, acqparams.txt, containing the following rows:

0 -1 0 0.0665
0 1 0 0.0665

Here a -1 means AP, and 1 means PA. The last column is the readout time. If you have any questions about this, ask your scanner technician. "What if he's kind of creepy?" I don't care, girl; you gotta find out somehow.

While you're at it, also ask which image from the scan was the AP scan, and which was the PA scan. Once you have identified each, you should combine the two using fslmerge:

fslmerge -t AP_PA_scan AP_scan PA_scan 

Once you have that, you have all you need for topup:

topup --imain=AP_PA_scan --datain=acqparams.txt --out=topup_AP_PA_scan

This will produce an image, topup_AP_PA_scan, of where the distortions are located in your DTI scan. This will then be called by the eddy tool to undo any distortions at the same time that it does eddy correction.

The only other preparation you need for the eddy correction is an index list of where the acquisition parameters hold (which should usually just be a 1 for each row for each scan), and a mask excluding any non-brain content, which you can do with bet:

bet dwi_image betted_dwi_image -m -f 0.2

Then feed this into the eddy command (note that a bvecs and bvals image should be automatically generated if you imported your data using a tool such as dcm2niigui):

eddy --imain=dwi_image --mask=betted_dwi_image --index=index.txt --acqp=acqparams.txt --bvecs=bvecs --bvals=bvals --fwhm=0 --topup=topup_AP_PA_scan --flm=quadratic --out=eddy_unwarped_images

Note that the --fwhm and --flm arguments I chose here are the defaults that FSL recommends.

The differences before and after can be seen in fslview:

Of course, the distortion correction step isn't strictly necessary; and you can still do a much simpler version of eddy correction by simply selecting FDT Diffusion from the FSL menu and then selecting the Eddy Correction tab, which only requires input and output arguments. All I'm saying is that, if you're feeling particularly litigious, you should probably be safe and do both steps.

Introduction to Diffusion Tensor Imaging: From Hospital Horror Story to Neuroimaging

It is well known that one of the deepest human fears is to be a patient in a hospital late at night, all alone, while a psychotic nurse injects you with a paralyzing agent, opens up your skull with a bone saw, and begins peeling away layers of your cortex while you are still awake.*

As horrifying as this nightmare scenario may be, it also lends an important insight into an increasingly popular neuroimaging method, diffusion tensor imaging (DTI; pronounced "diddy"). To be more gruesomely specific, the psychotic nurse is able to peel away strips of your brain because it has preferred tear directions - just like string cheese. These strips follow the general direction of fascicles, or bundles of nerves, comprising grey and white matter pathways; and of these pathways, it is white matter that tends to exhibit a curious phenomenon called anisotropy.

Imagine, for example, that I release a gas - such as, let's say, deadly neurotoxin - into a spherical compartment, such as a balloon. The gas, through a process called Brownian motion (not to be confused with the Dr. Will Brown frequently mentioned here) will begin to diffuse randomly in all directions; in other words, as though it is unconstrained.

However, release the same gas into a cylindrical or tube-shaped compartment, such as one of those cardboard tubes you used to fight with when you were a kid,** and the gas particles will tend to move along the direction of the tube. This is what is known as anisotropy, meaning that the direction of the diffusion tends to go in a particular direction, as opposed to isotropy, where diffusion occurs in all directions with equal probability.

Left two figures: Ellipsoids showing different amounts of anisotropy, with lambda 1, 2, and 3 symbolizing eigenvalues. Eigenvalues represent the amount of diffusion along a particular direction. Right: A sphere representing isotropy, where diffusion occurs with equal probability in all directions.

This diffusion can be measured in DTI scans, which in turn can be used to indirectly measure white matter integrity and structural connectivity between different areas of the brain. A common use of DTI is to compare different populations, such as young and old, and to observe where fractional anisotropy (FA) differs between groups, possibly with the assumption that less FA can be indicative of less efficient communication between cortical regions. There are other applications as well, but this is the one we will focus on for the remainder of the tutorials.

The data that we will be using can be found on the FSL course website, after scrolling down to Data Files and downloading File 2 (melodic and diffusion). I haven't been able to find any good online repositories for DTI data, so we'll be working with a relatively small sample of three subjects in one group, and three subjects in the other. Also note that while we will focus on FSL, there are many other tools that process DTI data, including some new commands in AFNI, and also a program called TORTOISE. As with most things I post about, these methods have already been covered in detail by others; and in particular I recommend a blog called blog.cogneurostats.com, which covers both the AFNI and TORTOISE approaches to DTI, along with other tutorials that I thought I had been the first to cover, but which have actually already been discussed in detail. I encourage you to check it out - but also to come back, eventually.

*Or maybe that's just me.
**And maybe you still do!