Using Subject Space ROIs from Freesurfer#

An example using the AFQ API to find bundles as defined by endpoint ROIs from freesurfer. This example can be modified to work with ROIs in subject space from pipelines other than freesurfer.

import os.path as op

import nibabel as nib
import plotly
import numpy as np

from AFQ.api.group import GroupAFQ
import AFQ.data.fetch as afd
from AFQ.definitions.image import RoiImage
import AFQ.api.bundle_dict as abd
2026-05-26 22:58:36,475	INFO util.py:154 -- Missing packages: ['ipywidgets']. Run `pip install -U ipywidgets`, then restart the notebook server for rich notebook output.

Get some example data#

Retrieves High angular resolution diffusion imaging (HARDI) dataset from Stanford’s Vista Lab

see https://purl.stanford.edu/ng782rw8378 for details on dataset.

The data for the first subject and first session are downloaded locally (by default into the users home directory) under:

.dipy/stanford_hardi/

Anatomical data (anat) and Diffusion-weighted imaging data (dwi) are then extracted, formatted to be BIDS compliant, and placed in the AFQ data directory (by default in the users home directory) under:

AFQ_data/stanford_hardi/

This data represents the required preprocessed diffusion data necessary for initializing the GroupAFQ object (which we will do next)

The clear_previous_afq is used to remove any previous runs of the afq object stored in the AFQ_data/stanford_hardi/ BIDS directory. Set it to None if you want to use the results of previous runs. Setting it to “track” as here will only clear derivatives that depend on the tractography stage (i.e., bundle delination and tract profile calculation), as well as the tractography itself, to save time on recomputation. If you want to only clear derivatives that depend on bundle delineation, and keep the tractography, you can set clear_previous_afq to “recog” instead.

afd.organize_stanford_data(clear_previous_afq="track")
---------------------------------------------------------------------------
KeyboardInterrupt                         Traceback (most recent call last)
Cell In[2], line 1
----> 1 afd.organize_stanford_data(clear_previous_afq="track")

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/site-packages/AFQ/data/fetch.py:1819, in organize_stanford_data(path, clear_previous_afq)
   1817 # fetches data for first subject and session
   1818 logger.info("fetching Stanford HARDI data")
-> 1819 dpd.fetch_stanford_hardi()
   1821 if path is None:
   1822     if not op.exists(afq_home):

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/site-packages/dipy/data/fetcher.py:494, in _make_fetcher.<locals>.fetcher(include_optional)
    491         continue
    492     files[str(n)] = (baseurl + f, md5_list[i] if md5_list is not None else None)
--> 494 fetch_data(files, folder, data_size=data_size, use_headers=use_headers)
    496 if msg is not None:
    497     logger.info(msg)

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/site-packages/dipy/testing/decorators.py:201, in warning_for_keywords.<locals>.decorator.<locals>.wrapper(*args, **kwargs)
    194 # Check if the current version is within the warning range
    195 if (
    196     version.parse(from_version)
    197     <= version.parse(current_version)
    198     <= version.parse(until_version)
    199 ):
    200     # Convert positional to keyword arguments and issue a warning
--> 201     return convert_positional_to_keyword(func, args, kwargs)
    203 # If the version is greater than the until_version,
    204 # pass the arguments as they are
    205 elif version.parse(current_version) > version.parse(until_version):

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/site-packages/dipy/testing/decorators.py:192, in warning_for_keywords.<locals>.decorator.<locals>.wrapper.<locals>.convert_positional_to_keyword(func, args, kwargs)
    182         warnings.warn(
    183             f"Pass {positionally_passed_kwonly_args} as keyword args. "
    184             f"From version {until_version} passing these as positional "
   (...)    187             stacklevel=3,
    188         )
    190     return func(*positional_args, **corrected_kwargs)
--> 192 return func(*args, **kwargs)

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/site-packages/dipy/data/fetcher.py:397, in fetch_data(files, folder, data_size, use_headers, raise_on_error)
    395 logger.info(f"From: {url}")
    396 try:
--> 397     _get_file_data(fullpath, url, use_headers=use_headers, stored_md5=md5)
    398     successful_downloads += 1
    399 except (FetcherError, Exception) as e:

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/site-packages/dipy/data/fetcher.py:262, in _get_file_data(fname, url, use_headers, timeout, max_retries, stored_md5)
    260 with open(fname, "wb") as data:
    261     if response_size is None:
--> 262         copyfileobj(opener, data)
    263     else:
    264         copyfileobj_withprogress(opener, data, response_size)

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/shutil.py:203, in copyfileobj(fsrc, fdst, length)
    201 fsrc_read = fsrc.read
    202 fdst_write = fdst.write
--> 203 while buf := fsrc_read(length):
    204     fdst_write(buf)

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/http/client.py:478, in HTTPResponse.read(self, amt)
    475     return b""
    477 if self.chunked:
--> 478     return self._read_chunked(amt)
    480 if amt is not None and amt >= 0:
    481     if self.length is not None and amt > self.length:
    482         # clip the read to the "end of response"

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/http/client.py:608, in HTTPResponse._read_chunked(self, amt)
    605     self.chunk_left = chunk_left - amt
    606     break
--> 608 value.append(self._safe_read(chunk_left))
    609 if amt is not None:
    610     amt -= chunk_left

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/http/client.py:648, in HTTPResponse._safe_read(self, amt)
    641 """Read the number of bytes requested.
    642 
    643 This function should be used when <amt> bytes "should" be present for
    644 reading. If the bytes are truly not available (due to EOF), then the
    645 IncompleteRead exception can be used to detect the problem.
    646 """
    647 cursize = min(amt, _MIN_READ_BUF_SIZE)
--> 648 data = self.fp.read(cursize)
    649 if len(data) >= amt:
    650     return data

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/socket.py:719, in SocketIO.readinto(self, b)
    717     raise OSError("cannot read from timed out object")
    718 try:
--> 719     return self._sock.recv_into(b)
    720 except timeout:
    721     self._timeout_occurred = True

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/ssl.py:1304, in SSLSocket.recv_into(self, buffer, nbytes, flags)
   1300     if flags != 0:
   1301         raise ValueError(
   1302           "non-zero flags not allowed in calls to recv_into() on %s" %
   1303           self.__class__)
-> 1304     return self.read(nbytes, buffer)
   1305 else:
   1306     return super().recv_into(buffer, nbytes, flags)

File /opt/hostedtoolcache/Python/3.13.13/x64/lib/python3.13/ssl.py:1138, in SSLSocket.read(self, len, buffer)
   1136 try:
   1137     if buffer is not None:
-> 1138         return self._sslobj.read(len, buffer)
   1139     else:
   1140         return self._sslobj.read(len)

KeyboardInterrupt: 

Generate left thalamus ROI from freesurfer segmentation file#

  1. Load the segmentation file that was generated by Freesurfer for the specific subject.

  2. Identify the left thalamus within the file, which has the label number 41

  3. Create a Nifti image representing the left thalamus ROI:

    • Assign a value of 1 to the voxels that Freesurfer has labeled as 41 (i.e., the left thalamus).

    • Assign a value of 0 to all other voxels. This binary mask format is the expected input for pyAFQ when dealing with subject space ROIs. If it’s already in binary format, there is no need to do this step.

freesurfer_subject_folder = op.join(
    afd.afq_home, "stanford_hardi",
    "derivatives", "freesurfer",
    "sub-01", "ses-01",
    "anat")

seg_file = nib.load(op.join(
    freesurfer_subject_folder, "sub-01_ses-01_seg.nii.gz"))
left_thal = seg_file.get_fdata() == 41
nib.save(
    nib.Nifti1Image(
        left_thal.astype(np.float32),
        seg_file.affine),
    op.join(
        freesurfer_subject_folder,
        "sub-01_ses-01_desc-leftThal_mask.nii.gz"))

# Fetch LV1 ROI
# which was already generated using the process above
afd.fetch_stanford_hardi_lv1()

Set tractography parameters (optional)#

We make this tracking_params which we will pass to the GroupAFQ object which specifies that we want 10,000 seeds randomly distributed only within the endpoint ROIs and not throughout the white matter. This is controlled by passing "seed_mask": RoiImage() in the tracking_params dict.

We only do this to make this example faster and consume less space.

tracking_params = dict(n_seeds=10000,
                       random_seeds=True,
                       rng_seed=42,
                       seed_mask=RoiImage(use_endpoints=True))

Define custom BundleDict object#

In a typical BundleDict object, ROIs are passed as paths to Nifti files. Here, we define ROIs as dictionaries instead, containing BIDS filters. Then pyAFQ can find the respective ROI for each subject and session.

bundles = abd.BundleDict({
    "L_OR": {
        "start": {
            "scope": "freesurfer",
            "suffix": "mask",
            "desc": "leftThal"},
        "end": {
            "scope": "freesurfer",
            "suffix": "anat",
            "desc": "LV1"
        },
        "cross_midline": False,
        "space": "subject"
    }})

Initialize a GroupAFQ object:#

Creates a GroupAFQ object, that encapsulates tractometry, passing in our custom bundle info. Then we run the pipeline and generate a visualization of the bundle we found.

myafq = GroupAFQ(
    bids_path=op.join(afd.afq_home, 'stanford_hardi'),
    dwi_preproc_pipeline='vistasoft',
    t1_preproc_pipeline='freesurfer',
    tracking_params=tracking_params,
    bundle_info=bundles)

bundle_html = myafq.export("indiv_bundles_figures")
plotly.io.show(bundle_html["01"]["L_OR"])