# Pipeline tutorial¶

## 0. Before you begin¶

To get started, first [create a singularity image of xcpEngine:

$singularity build /data/applications/xcpEngine.simg docker://pennbbl/xcpengine:latest  Next, download an example output from FMRIPREP Suppose the downloaded data is extracted to ${DATA_ROOT}/fmriprep, where ${DATA_ROOT} is an existing directory on your system. ## 1. Running the anatomical pipeline¶ You need to create a cohort file and a design file to show XCP where your data is and tell it what to do with it. Since there is only a single subject with a single session, write the following content to${DATA_ROOT}/anat_cohort.csv:

id0,img
sub-1,fmriprep/sub-1/anat/sub-1_T1w_preproc.nii.gz


Then download the antsCT design file into ${DATA_ROOT}/anat-antsct.dsn Now you’re ready to run the anatomical pipeline! Create an empty directory in your home directory (we’ll use ${HOME} because it is directly mounted by Singularity by default, so do mkdir ${HOME}/data). This will be the bind point for your system’s ${DATA_ROOT} directory and will let singularity access your files. Binding directories is very important and worth understanding well. See the singularity documentation for details.:

singularity run -B ${DATA_ROOT}:${HOME}/data  \
/data/applications/xcpEngine.simg
-d ${HOME}/data/anat-antsct.dsn \ -c${HOME}/data/anat_cohort.csv  \
-o ${HOME}/data/xcp_output \ -t 1 \ -r${HOME}/data


This will take up to 2 days, but when it’s done you will have the full output of antsCorticalThickness for this subject! It also provides all the spatial normalization info to map atlases to your BOLD data. See the Anatomical processing streams stream to learn about the available templates and atlases.

## 2. Running a functional connectivity pipeline¶

Processing fMRI data for functional connectivity analysis can be done using another design file. Now save one of the nicely-performing pipeline design files as something like ${DATA_ROOT}/fc-36p.dsn. This will take the preprocessed BOLD output from FMRIPREP and prepare it for functional connectivity analysis. Create a new cohort csv that tells XCP where the output from the struc module is located and where the output from FMRIPREP is located. In ${DATA_ROOT}/func_cohort.csv write:

id0,antsct,img


This specifies that we will process the task-rest scan from this subject. Other runs from the same session would need to be added as additional lines in the cohort file. Run xcpEngine with this new cohort file:

singularity run -B ${DATA_ROOT}:${HOME}/data  \
/data/applications/xcpEngine.simg
-d ${HOME}/data/fc-36p.dsn \ -c${HOME}/data/func_cohort.csv  \
-o ${HOME}/data/xcp_output \ -t 1 \ -r${HOME}/data


## 3. Running a ASL processing pipeline¶

Processing ASL data for computation of CBF can be done using design files This will take the ASL, M0 (if available) and anatomical directory. ASL processing steps is the same as both anatomical and functional connectivity pipelines, the only different is cohort file and design files.

## 4. Arguments¶

While the pipeline is running, let’s break down the call that we made to the XCP Engine. We passed a total of 5 arguments to the pipeline.

• A design file, -d ${HOME}/data/example/anat-antsct.dsn • A cohort file, -c${HOME}/data/example/anat_cohort.csv
• An output path, ${HOME}/data/example/xcp_output • A verbosity level, -t 1 • A reference/relative directory, -r${HOME}/data

Let’s discuss each of these, and how a user might go about selecting or preparing them.

### Design file¶

The design file parametrizes the image processing stream. The XCP system supports multimodal image processing, including inter alia functional connectivity and volumetric anatomy. How does the system know which of these available processing streams it should execute? It parses the parameters provided by the user in the design file. In our example, we parametrized the stream using the design file anat-antsct.dsn.

Near the top of the file, you will find a variable called pipeline that should look something like: confound,regress,fcon,reho,alff,net,roiquant,seed,norm,qcfc. The pipeline variable tells the XCP system which modules it should run, and what order they should be run in.

Underneath the pipeline variable, you will find code blocks corresponding to each of the modules defined in pipeline. If you’re curious as to what effects any of the variables have, just look up the variables in the documentation for the relevant pipeline Modules.

### Cohort file and reference directory¶

The design file instructs the pipeline as to how inputs should be processed, but the cohort file (also called a subject list) actually informs the pipeline where to find the inputs. Let’s look at the cohort file that we used for this analysis.:

id0,antsct,img


The cohort file is formatted as a .csv with 3 variables and 1 observation (subject). The first line of the cohort file is a header that defines each of the variables. Subject identifiers are placed in columns starting with id and ending with a non-negative integer. For instance, the first identifier (id0) of the first subject is sub-1. There could be a second identifier (id1) such as ses-01 if needed.

The inputs for each subject are defined in the remaining columns, here antsct and fmriprep. antsct defines the path to the output files of the subject’s processed ANTs Cortical Thickness pipeline (which has already been run as part of the Anatomical processing streams stream in step 1). fmriprep defines the prefix to the main image that this pipeline will analyze. Since this is the cohort for a functional connectivity stream, the main image will be a functional image (in this case, resting state).

If we look at our call to xcpEngine, we can see that we passed it the argument -r ${DATADIR}. This argument instructs xcpEngine to search within ${DATADIR} for cohort paths. This is very useful when using Singularity of Docker, as you can specify the relative bind path as your root while keeping the paths in your cohort file relative to your system’s root.

Now, let’s suppose that we have already processed this subject through the pipeline system, and we acquire data for a new, 2nd subject. Let’s say this new subject has identifier sub-2. To process this new subject, DO NOT CREATE A NEW COHORT FILE. Instead, edit your existing cohort file and add the new subject as a new line at the end of the file. For our example subject, the corresponding line in the cohort file might be something like sub-2,xcp_output/sub-2/struc,fmriprep/sub-2/func/sub-2_task-rest_space-T1w_desc-preproc_bold.nii.gz. Why edit the existing cohort file instead of creating a new one?

• The pipeline will automatically detect that it has already run for the other subject, so it will not waste computational resources on them.
• The pipeline will then collate group-level data across all 8 subjects. If you were to create a new cohort file with just the new subject, group-level data would be pulled from only that subject. Not much of a group, then.

## 5. Output files¶

To see what the remaining arguments to xcpEngine do, we will need to look at the pipeline’s output. By now, the pipeline that you launched earlier will hopefully have executed to completion. Let’s take a look at the output directory that you defined using the -o option, ${output_root}. If you list the contents of ${output_root}, you will find 7 subject-level output directories (corresponding to the values of the id0 variable in the cohort file) and one group-level output directory (called group). (You can change the group-level output path using the additional command-line argument -a out_group=<where you want the group-level output>.)

Begin by looking at the subject-level output. Navigate to the first subject’s output directory, ${output_root}/sub-1. In this directory, you will find: • A subject-specific copy of the design file that you used to run the pipeline, evaluated and modified to correspond to this particular subject (sub-1). (In the XCP system, the process of mapping the template design file to each subject is called localisation, and the script that handles this is called the localiser.) • An atlas directory (sub-1_atlas). Inside the atlas directory, each parcellation that has been analyzed will exist as a NIfTI file, registered to the subject’s T1w native space. • A subdirectory corresponding to each pipeline module, as defined in the pipeline variable in the design file. For the most part, these directories store images and files that the pipeline uses to verify successful processing. • Take a look inside the fcon subdirectory. Inside, there will be a separate subdirectory for each of the atlases that the pipeline has processed. For instance, in the power264 subdirectory (corresponding to the 264-node Power atlas), there will be files suffixed ts.1D and network.txt. • ts.1D contains 264 columns corresponding to each node of the atlas; each column contains a region’s functional time series. • network.txt contains the functional connectivity matrix or connectome for the Power atlas, formatted as a vector to remove redundant edges. • A log directory (sub-1_logs). Inside the log directory, open the file whose name ends with _LOG. This is where all of the pipeline’s image processing commands are logged. The verbosity of this log can be modified using the argument to the -t option). It is recommended that you use a verbosity level of either 1 or 2. For most cases, 1 will be sufficient, but 2 can sometimes provide additional, lower-level diagnostic information. • A quality file (sub-1_quality.csv). The contents of the quality file will be discussed in detail later, along with group-level outputs. • A spatial metadata file (sub-1_spaces.json). The pipeline uses this to determine how to move images between different coordinate spaces. • The final output of processing (sub-1.nii.gz). This is the primary functional image, after all image processing steps (except for smoothing) have been applied to it. If you have smoothing in your design file, smoothed outputs are saved separately as files like sub-1_img_sm${k}.nii.gz inside the norm and regress folders, with ${k} the smoothing kernel size. However, this preprocessed file usually isn’t as useful for analysis as are its derivatives, which brings us to … • An index of derivative images (sub-1_derivatives.json). • Let’s look at the content of the derivatives file now. Run the command shown, and find the entry for reho. This JSON object corresponds to the voxelwise map of this subject’s regional homogeneity (ReHo). • The map can be found in the path next to the Map attribute. (You can open this in fslview if you would like.) • The Provenance attributes tell us that the map was produced as part of the 6th pipeline module, reho. • The Space attribute tells us that the map is in 2mm isotropic MNI space. • The Statistic attribute instructs the pipeline’s roiquant module that it should compute the mean value within each parcel of each atlas when converting the voxelwise derivative into an ROI-wise derivative. • The Type attribute is used by the pipeline when it makes decisions regarding interpolations and other processing steps. • There will actually be a separate index for each coordinate space that has been processed. Note that there’s also a sub-1_derivatives-sub-1_fc.json, which has the same metadata for derivatives in the subject’s native functional space. Next, let’s examine the group-level output. Navigate to ${output_root}/group. In this directory, you will find:

• The dependency metadata from earlier (dependencies/*pipelineDescription.json). (A new time-stamped metadata file is generated for each run of the pipeline.)
• An error logging directory (error). This should hopefully be empty!
• A log directory (log), analogous to the log directory from the subject level.
• Module-level directories, in this case for the roiquant and qcfc modules.
• Let’s look at the group-level roiquant output. Like the subject-level net output, there will be a separate subdirectory for each atlas that has been processed.
• Inside the atlas-level subdirectory, there will be files corresponding to any derivatives that had a non-null value for their Statistic attribute. For instance, the ReHo that we looked at earlier (Statistic: mean) has been quantified regionally and collated across all subjects in the file ending with the suffix RegionalMeanReho.csv. You may wish to examine one of these files; they are ready to be loaded into R or any other environment capable of parsing .csv s.
• A sample quality file for the modality (fc_quality.csv).
• The qcfc module’s subdirectory will contain reports analogous to those from our .These aren’t really useful for a sample of only 1 subject, so we won’t look at them here.
• Collated subject-level quality indices (n1_quality.csv, not to be confused with the sample-level quality file). If you examine this file, you will find the quality indices that the functional connectivity stream tracks. This file can be used to establish exclusion criteria when building a final sample, for instance on the basis of subject movement or registration quality.
• An audit file (n1_audit.csv). This file indicates whether each pipeline module has successfully run for each subject. 1 indicates successful completion, while 0 indicates a nonstandard exit condition.

## 6. Anatomy of the pipeline system¶

Now, let’s pull this information together to consider how the pipeline system operates.

1. The front end, xcpEngine, parses the provided design and cohort files.
2. The localiser uses the information in the cohort file to generate a subject-specific version of the design file for each subject. (The localiser shifts processing from the sample level to the subject level; this is called the localisation or map step.)
3. xcpEngine parses the pipeline variable in the design file to determine what Modules (or processing routines) it should run. Different imaging and data modalities (e.g., anatomical, functional connectivity, task activation) will make use of a different series of modules.
4. xcpEngine submits a copy of each module for each subject in the cohort using that subject’s local design file. Modules run in series, with all subjects running each module in parallel. As it runs, each module writes derivatives and metadata to its output directory.
5. To collate subject-level data or perform group-level analysis, the pipeline uses the delocaliser. Shift of processing from the subject level to the sample level is called delocalisation or a reduce step.

## 7. Getting help¶

To get help, the correct channel to use is  Github <https://github.com/PennBBL/xcpEngine/issues>_. Open a new issue and describe your problem. If the problem is highly dataset-specific, you can contact the development team by email, but Github is almost always the preferred channel for communicating about pipeline functionality. You can also use the issue system to request new pipeline features or suggest changes.

## 8. Common Errors¶

A non-exhaustive list of some common errors, and fixes to try.

• ImportError: bad magic number in 'site' : Try running unset PYTHONPATH immediately prior to running the pipeline.
• Cannot allocate vector of size xx Mb` : Try increasing the amount of memory available for running the pipeline.