Research organism

elife

eLife

2050-084X

eLife Sciences Publications, Ltd

59430

10.7554/eLife.59430

Research Article

Neuroscience

Multi-contrast anatomical subcortical structures parcellation

Bazin

Pierre-Louis

https://orcid.org/0000-0002-0141-5510

pilou.bazin@uva.nl12Alkemade

Anneke

http://orcid.org/0000-0002-3234-353X

1Mulder

Martijn J

13Henry

Amanda G

http://orcid.org/0000-0002-2923-4199

4Forstmann

Birte U

http://orcid.org/0000-0002-1005-1675

11Integrative Model-based Cognitive Neuroscience research unit, University of Amsterdam

Amsterdam

Netherlands2Max-Planck Institute for Human Cognitive and Brain Sciences

Leipzig

Germany3Psychology Department, Utrecht University

Utrecht

Netherlands4Faculty of Archaeology, Leiden University

Leiden

Netherlands

Verstynen

Timothy

Reviewing EditorCarnegie Mellon UniversityUnited StatesFrank

Michael J

Senior EditorBrown UniversityUnited States

16122020

2020

e59430

2805202015122020

2020

Bazin et al

http://creativecommons.org/licenses/by/4.0/

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

The human subcortex is comprised of more than 450 individual nuclei which lie deep in the brain. Due to their small size and close proximity, up until now only 7% have been depicted in standard MRI atlases. Thus, the human subcortex can largely be considered as terra incognita. Here, we present a new open-source parcellation algorithm to automatically map the subcortex. The new algorithm has been tested on 17 prominent subcortical structures based on a large quantitative MRI dataset at 7 Tesla. It has been carefully validated against expert human raters and previous methods, and can easily be extended to other subcortical structures and applied to any quantitative MRI dataset. In sum, we hope this novel parcellation algorithm will facilitate functional and structural neuroimaging research into small subcortical nuclei and help to chart terra incognita.

subcortexanatomical parcellationquantitative MRI

Research organismHuman

http://dx.doi.org/10.13039/501100003246

Nederlandse Organisatie voor Wetenschappelijk Onderzoek

VICI

Forstmann

Birte U

http://dx.doi.org/10.13039/501100003246

Nederlandse Organisatie voor Wetenschappelijk Onderzoek

STW

Alkemade

Anneke

Forstmann

Birte U

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Author impact statement

An open-source software tool enables the anatomical parcellation of an unprecedented number of subcortical structures in magnetic resonance images of the human brain, automatically and in individual subjects.

Introduction

Subcortical brain structures are often neglected in neuroimaging studies due to their small size, limited inter-regional contrast, and weak signal-to-noise ratio in functional imaging (Forstmann et al., 2016; Johansen-Berg, 2013). Yet, these small and diverse structures are prominent nodes in functional networks (Marquand et al., 2017; Ji et al., 2019), and they undergo pathological alterations already at early stages of neurodegenerative diseases (Andersen et al., 2014; Koshiyama et al., 2018). Deep brain stimulation surgery, originally performed to reduce motor symptoms in essential tremors, is now a promising therapeutic option in later stages of Parkinson’s disease and movement disorders, as well as refractory psychiatric illnesses in obsessive-compulsive disorder, anorexia, or depression (Forstmann et al., 2017; Mosley et al., 2018). Evolutionary genetics even uncovered that in modern humans, Neanderthal-inherited alleles were preferentially down-regulated in subcortical and cerebellar regions compared to other brain regions (McCoy et al., 2017), suggesting these structures to be essential in making us specifically human.

Despite their importance, these areas are particularly difficult to image. Furthermore, the size, shape, and location of these brain regions changes with development and aging (Fjell et al., 2013; Keuken et al., 2013; Yeatman et al., 2014; Herting et al., 2018). Experience-based plasticity continuously remodels myelin (Tardif et al., 2016; Hill et al., 2018; Turner, 2019), iron and other magnetic substances accumulate with age or pathology (Andersen et al., 2014; Zhang et al., 2018), both bringing changes in the MRI appearance of subcortical regions with diverse tissue characteristics (Draganski et al., 2011; Keuken et al., 2017).

Thus, mapping the structure and function of the subcortex is a major endeavor as well as a major challenge for human neuroscience. Extensive work available from animal brain models unfortunately does not translate in a straightforward way to human subcortical anatomy nor does it shed much light on its involvement in human cognition (Steiner and Tseng, 2017). Besides serious difficulties in obtaining adequate measures of subcortical neural activity in functional MRI (de Hollander et al., 2017; Miletić et al., 2020), atlases and techniques for labeling accurately and reliably individual subcortical structures have also been scarce (Frazier et al., 2005; Chakravarty et al., 2006; Ahsan et al., 2007; Yelnik et al., 2007; Qiu et al., 2010; Patenaude et al., 2011), typically labeling the thalamus, striatum (or its subdivision into caudate and putamen), and globus pallidus (internal and external segments combined), sometimes the amygdala. However, recent advances in anatomical MRI, combining multiple contrasts and/or quantitative MRI mapping and utilizing the higher resolution achievable with 7 Tesla (7T) and above have started to reduce the gap, each mapping a few additional structures or sub-structures, primarily the iron-rich substantia nigra, red nucleus and sub-thalamic nucleus (Keuken et al., 2013; Xiao et al., 2015; Visser et al., 2016a; Visser et al., 2016b; Wang et al., 2016; Makowski et al., 2018; Ewert et al., 2018; Iglesias et al., 2018; Pauli et al., 2018; Sitek et al., 2019). While these efforts generated valuable atlases, they do not yet enable to identify many subcortical structures in individual subjects. Manual delineation, on the other hand, requires extensive manual labor from highly trained experts which cannot be easily applied to large cohorts or clinical settings.

Here, we propose a new automated parcellation technique to identify and label 17 individual subcortical structures of varying size and composition in individual subjects, based on a large quantitative 7T MRI database (Alkemade et al., 2020), using quantitative maps of relaxation rates R1 and R2* (1/T1 and 1/T2*, respectively) and quantitative susceptibility maps (QSM) as anatomical contrasts. The algorithm, named Multi-contrast Anatomical Subcortical Structure Parcellation (MASSP), follows a Bayesian multi-object approach similar in essence to previous efforts (Fischl et al., 2002; Eugenio Iglesias et al., 2013; Visser et al., 2016a; Garzón et al., 2018), combining shape priors, intensity distribution models, spatial relationships, and global constraints. The main innovation of our approach is to explicitly estimate interfaces between subcortical structures based on a joint model derived from signed distance functions. Modeling interfaces in addition to the structure itself provides a rich basis to encode relationships and anatomical knowledge in shape and intensity priors. A voxel-wise Markovian diffusion regularizes the combined priors for each defined interface, lowering the imaging noise. Finally, the voxel-wise posteriors for the different structures and interfaces are further combined into global anatomical parcels by topology correction and region growing taking into account volumetric priors, which regularizes parcellation results further in smaller nuclei with low or heterogeneous contrast. To validate the results from this new method, in a thorough comparison with expert manual labeling, we show that the proposed method provides results very close from manual raters in many structures and exhibit reasonable bias across the adult lifespan. The method can easily be extended to new structures, can be applied to any quantitative MRI dataset and is available in Open Source as part of Nighres (Huntenburg et al., 2018), a neuroimage analysis package aimed at high-resolution neuroimaging.

Results

The MASSP parcellation method presented here has been trained to parcellate the following 17 structures: striatum (Str), thalamus (Tha), lateral, 3rd and 4th ventricles (LV, 3V, 4V), amygdala (Amg), globus pallidus internal segment (GPi) and external segment (GPe), SN, STN, red nucleus (RN), ventral tegmental area (VTA), fornix (fx), internal capsule (ic), periaqueductal gray (PAG), pedunculopontine nucleus (PPN), and claustrum (Cl), see Figure 1. These structures include the most commonly defined subcortical regions (Str, Tha, Amg, LV), the main iron-rich nuclei (GPi, GPe, RN, SN, STN), as well as smaller, less studied areas (VTA, PAG, PPN, Cl), white matter structures (ic, fx), and the central ventricles (3V, 4V).

Figure 1.The 17 subcortical structures currently included in the parcellation algorithm in axial (<bold>A</bold>), sagittal (<bold>B</bold>), and coronal (<bold>C</bold>) views.

MASSP uses a data set of ten expert delineations as a basis for its modeling. From the delineations, an atlas of interfaces between structures, shape skeletons, and interface intensity histograms are generated, and used as prior in a multiple-step non-iterative Bayesian algorithm, see Figure 2 and Materials and methods.

Figure 2.The MASSP parcellation pipeline.

Atlas priors for interfaces between structures are combined to the MRI data, regularized via probability diffusion and topology correction, and the final structure posteriors are jointly estimated by region growing.

Validation against manual delineations

In a leave-one-out validation study comparing performance with the manual delineations, MASSP performed above 95% of the level of quality of the raters for Str, Tha, 4V, GPe, SN, RN, VTA, ic in terms of Dice overlap, the most stringent of the quality measures (see Figures 3 and 4 and Table 1). Several of the smaller structures have lower overlap ratios likely due to their smaller size (GPi, STN, PAG, PPN). Structures with an elongated shape (fx, Cl) remain challenging, due to the fact that small differences in location can substantially reduce overlap (Bazin et al., 2016). Despite these challenges, when comparing the dilated Dice scores, all structures were above 75% of overlap, with most reaching over 90% of the manual raters ability. Note that the Dice coefficient is very sensitive to size, as smaller structures will have lower overlap ratios for the same number of misclassified voxels. The dilated Dice coefficient is more representative of the variability regardless of size, as the smaller structures can reach high levels of overlap, both in manual and automated parcellations (see Table 1). The average surface distance confirms these results, showing values generally between one and two voxels of distance at a resolution of 0.7 mm, except in the cases of Amg, LV, fx, PPN, and Cl. These structures are generally more variable (LV), elongated (fx, Cl), or have a particularly low contrast with neighboring regions (Amg, PPN).

Table 1.Mean overlap and distance measures for the leave-one-out validation.

	Str	STN	SN	RN	GPi	GPe	Tha	LV	3V	4V	Amg	ic	VTA	fx	PAG	PPN	Cl
Dice overlap
MASSP	0.893	0.648	0.805	0.870	0.702	0.800	0.867	0.849	0.741	0.869	0.723	0.745	0.570	0.527	0.641	0.496	0.485
Manual	0.897	0.800	0.841	0.875	0.762	0.813	0.877	0.907	0.797	0.882	0.866	0.732	0.574	0.823	0.791	0.665	0.727
Ratio	0.995	0.811	0.957	0.996	0.925	0.987	0.989	0.936	0.936	0.988	0.836	1.020	0.994	0.641	0.814	0.754	0.664
Dilated overlap
MASSP	0.982	0.919	0.977	0.991	0.909	0.956	0.970	0.929	0.890	0.951	0.891	0.915	0.863	0.756	0.897	0.795	0.789
Manual	0.987	0.988	0.985	0.995	0.953	0.972	0.970	0.967	0.944	0.961	0.978	0.924	0.818	0.957	0.960	0.910	0.914
Ratio	0.995	0.930	0.992	0.995	0.955	0.984	1.000	0.961	0.946	0.991	0.911	0.992	1.059	0.790	0.935	0.879	0.863
Average surface distance
MASSP	0.750	0.911	0.676	0.491	1.140	0.863	1.058	2.690	0.994	0.817	1.476	1.275	1.074	2.950	0.955	1.484	1.685
Manual	0.723	0.508	0.571	0.482	0.902	0.804	0.971	0.615	0.637	0.671	0.779	1.045	1.204	0.703	0.555	0.801	0.670
Ratio	0.971	0.590	0.852	0.996	0.861	0.943	0.916	0.277	0.662	1.020	0.553	0.834	1.161	0.287	0.610	0.619	0.465

Figure 3.Leave-one-out validation of the structures parcellated by MASSP, compared to the human rater with most neuroanatomical expertise.

Scores for the left and right side are computed separately and then combined into box-and-whisker plots.

Figure 4.Inter-rater variability for the human expert raters.

Scores for the left and right side are computed separately and then combined into box-and-whisker plots.

Comparison to other automated methods

To provide a basis for comparison, we applied other freely available methods for subcortical structure parcellation to the same 10 subjects. MASSP performs similarly to or better than Freesurfer, FSL FIRST and a multi-atlas registration using ANTs (see Table 2). Multi-atlas registration provides high accuracy in most structures as well, but is biased toward under-estimating the size of smaller and elongated structures where overlap is systematically reduced across the individual atlas subjects. Multi-atlas registration is also quite computationally intensive when using multiple contrasts at high resolution. Finally, MASSP provides many more structures than Freesurfer and FSL FIRST, and can be easily applied to new structures based on additional manual delineations.

Table 2.Comparison with multi-atlas registration, Freesurfer, and FSL FIRST.

	Str	STN	SN	RN	GPi	GPe	Tha	LV	3V	4V	Amg	Ic	VTA	Fx	PAG	PPN	Cl
Dice overlap
MASSP	0.893	0.648	0.805	0.870	0.702	0.800	0.867	0.849	0.741	0.869	0.723	0.745	0.570	0.527	0.641	0.496	0.485
Multi-atlas	0.855	0.662	0.760	0.820	0.742	0.796	0.859	0.734	0.660	0.691	0.761	0.718	0.626	0.478	0.674	0.539	0.398
Freesurfer	0.876				0.778		0.838	0.858	0.430	0.769	0.692
FSL FIRST	0.875				0.813		0.839				0.653
Dilated overlap
MASSP	0.982	0.919	0.977	0.991	0.909	0.956	0.970	0.929	0.890	0.951	0.891	0.915	0.863	0.756	0.897	0.795	0.789
Multi-atlas	0.976	0.938	0.968	0.989	0.947	0.968	0.970	0.920	0.920	0.908	0.921	0.939	0.924	0.798	0.943	0.871	0.811
Freesurfer	0.975				0.922		0.946	0.974	0.562	0.911	0.857
FSL FIRST	0.976				0.946		0.950				0.843
Average surface distance
MASSP	0.750	0.911	0.676	0.491	1.140	0.863	1.058	2.690	0.994	0.817	1.476	1.275	1.074	2.950	0.955	1.484	1.685
Multi-atlas	0.961	0.891	0.858	0.675	0.992	0.882	1.083	1.417	0.932	1.249	1.359	1.129	0.813	1.362	0.794	1.055	1.273
Freesurfer	0.770				1.211		1.405	0.685	4.071	1.361	1.749
FSL FIRST	0.867				1.143		1.675				1.746
Volume bias
MASSP	0.041	0.017	-0.038	0.007	0.066	0.089	0.040	0.0470	0.121	0.047	0.078	0.183	0.032	-0.016	0.026	0.009	0.023
Multi-atlas	0.020	-0.087	-0.009	0.031	0.009	0.014	0.020	-0.003	-0.007	-0.092	-0.038	0.055	-0.067	-0.264	-0.090	-0.269	-0.376
Freesurfer	0.017				0.087		0.163	0.122	-0.551	0.351	0.468
FSL FIRST	-0.100				-0.021		0.165				-0.249

Application to new MRI contrasts

Quantitative MRI has only become recently applicable in larger studies, thanks in part to the development of integrated multi-parameter sequences (Weiskopf et al., 2013; Caan et al., 2019). Many data sets, including large-scale open databases, use more common T1- and T2-weighted MRI. In order to test the applicability of MASSP to such contrasts, we obtained the test-retest subset of the Human Connectome Project (HCP, Van Essen et al., 2013) and applied MASSP to the 45 pre-processed and skull-stripped T1- and T2-weighted images from each of the two test and retest sessions. While performing manual delineations on the new contrasts would be preferable, the model is already rich enough to provide stable parcellations. Test-retest reproducibility is similarly high for MASSP and Freesurfer, and are generally in agreement, see Figure 5 and Table 3.

Table 3.Test-retest comparison with Freesurfer on Human Connectome Project data.

	Str	STN	SN	RN	GPi	GPe	Tha	LV	3V	4V	Amg	ic	VTA	fx	PAG	PPN	Cl
Dice overlap
MASSP test-retest	0.914	0.701	0.818	0.829	0.791	0.859	0.928	0.881	0.837	0.870	0.866	0.860	0.738	0.774	0.714	0.713	0.785
Freesurfer test-retest	0.898				0.770		0.919	0.894	0.842	0.849	0.852
MASSP – Freesurfer	0.876				0.778		0.838	0.858	0.430	0.769	0.692
Dilated overlap
MASSP test-retest	0.987	0.939	0.977	0.978	0.963	0.977	0.990	0.980	0.979	0.986	0.981	0.973	0.969	0.961	0.965	0.972	0.966
Freesurfer test-retest	0.986				0.926		0.986	0.989	0.972	0.975	0.978
MASSP – Freesurfer	0.954				0.788		0.919	0.934	0.435	0.901	0.866
Average surface distance
MASSP test-retest	0.513	0.528	0.467	0.461	0.532	0.508	0.488	0.509	0.391	0.419	0.533	0.536	0.431	0.464	0.428	0.402	0.433
Freesurfer test-retest	0.876				0.778		0.838	0.858	0.430	0.769	0.692
MASSP – Freesurfer	0.976				1.673		1.605	1.946	5.699	1.428	1.478

Figure 5.Parcellation with Freesurfer (top, on T1w image) and MASSP (bottom, on T2w image) on Human Connectome Project data.

MASSP priors were not derived from the contrasts, but transferred via a spatial mapping of the quantitative MRI intensities from AHEAD subjects.

Biases due to atlas size

A common concern of brain parcellation methods is the risk of biases, as they are typically built from a small number of manual delineations. Our data set is part of a large scale study of the subcortex, for which we obtained manual delineations of the STN, SN, RN, GPe, and GPi on 105 subjects over the adult lifespan (18–80 year old, see Alkemade et al., 2020 for details). First, we investigated the impact of atlas size. We randomly assigned half of the subjects from each decade to two groups, and built atlas priors from subsets of 3, 5, 8, 10, 12, 15, and 18 subjects from the first group. The subjects used in the atlas were taken randomly from each decade (18-30, 31-40, 41-50,51-60, 61-70, 71-80), so as to maximize the age range represented in each atlas. Atlases of increasing size were constructed by adding subjects to previous atlases, so that atlases of increasing complexity include all subjects from simpler atlases. Results applying these atlases to parcellate the second group are given in Figure 6. As in previous studies (Eugenio Iglesias et al., 2013; Bazin and Pham, 2008), performance quickly stabilized with atlases of more than five subjects (no significant difference in Welch’s t-tests between using 18 subjects or any subset of 8 or more for all structures and measures).

Figure 6.MASSP parcellation scores as a function of increasing number of subjects included in the atlas.

Scores for the left and right side are computed separately and then combined into box-and-whisker plots.

Biases due to age differences

To more specifically test the influence of age on parcellation accuracy, we defined again six age groups by decade and randomly selected 10 subjects from each group. Each set of subjects was used as priors for the five structures above, and applied to the other age groups. Results are summarized in Figure 7. Examining this age bias, we can see a decrease in performance when parcellating subjects in the range of 60 to 80 years of age. The choice of priors seem to have a limited impact, which varies across structures. In particular, using priors from a similar age group is not always beneficial.

Figure 7.MASSP parcellation scores over the lifespan.

Each matrix show the average Dice overlap (top), dilated Dice overlap (middle), and average surface distance (bottom) for using one age group as prior (’train’) to parcellate another age group (’test’).

Bias on individual measures

Finally, we investigated the impact of this decrease in performance in the estimation of anatomical quantities, see Figure 8. The bias did affect the morphometric measures of structure volume and thickness, but the effects on the local measure of thickness was reduced compared to the global measure of volume. Quantitative MRI averages were very stable even when age biases are present in the parcellations.

Figure 8.Regression of volume (log scale), structure thickness, R1, R2*, and QSM MRI parameters estimated using manual delineations versus MASSP automated parcellations.

Circles show individual data points, linear regression is indicated by a straight line, and 95% confidence interval is given as the shaded area. Pearson correlation coefficients are indicated when significant (p-value<0.01).

For reference, we report structure volumes, thickness, R1, R2* and QSM values estimated from the entire AHEAD cohort for different age groups, extending our previous work based on manual delineations on a different data set (Keuken et al., 2017; Forstmann et al., 2014). Results are given in Table 4, describing average volumes, thickness, and quantitative MRI parameters for young, middle-aged, and older subjects for the 17 subcortical structures.

Table 4.Mean volume and quantitative MRI values for each age group.

Age	Str	STN	SN	RN	GPi	GPe	Tha	LV	3V	4V	Amg	ic	VTA	fx	PAG	PPN	Cl
Volume (m⁢m3)
18-40	10656	118	566	253	567	1366	7112	7524	1895	1391	1315	4335	254	1632	250	193	843
41-60	10572	124	583	256	586	1403	7492	8850	2024	1408	1363	4495	264	1808	255	195	830
61-80	10734	130	584	260	586	1397	7463	9142	2023	1407	1321	4407	272	1910	259	192	829
Thickness (m⁢m)
18-40	5.94	1.89	2.55	4.64	3.09	3.56	8.31	4.27	2.77	4.03	4.81	4.06	1.69	2.06	1.78	1.92	1.79
41-60	5.47	1.86	2.66	4.58	2.96	3.41	8.28	5.08	2.95	3.89	4.85	4.19	1.76	1.96	1.84	1.86	1.80
61-80	5.22	1.83	2.60	4.11	2.92	3.22	8.28	4.90	3.18	4.06	4.73	4.19	1.80	1.97	1.90	1.95	1.82
qR1 (H⁢z)
18-40	0.647	0.949	0.857	0.928	0.868	0.850	0.761	0.332	0.346	0.274	0.546	0.906	0.819	0.714	0.654	0.779	0.650
41-60	0.662	0.968	0.893	0.939	0.879	0.856	0.758	0.278	0.315	0.269	0.559	0.904	0.833	0.671	0.653	0.771	0.664
61-80	0.648	0.952	0.882	0.903	0.860	0.830	0.743	0.273	0.300	0.270	0.552	0.884	0.814	0.638	0.647	0.764	0.669
qR2* (H⁢z)
18-40	43.8	67.1	67.8	63.2	75.2	79.6	38.1	14.7	18.9	9.0	25.5	36.8	39.2	37.4	25.9	32.7	32.6
41-60	50.4	74.1	74.1	77.1	80.2	87.9	40.3	8.4	12.4	11.7	28.1	38.7	42.8	37.4	28.0	33.4	36.9
61-80	51.8	77.0	72.5	73.8	77.8	87.0	40.1	8.5	10.2	12.0	30.1	39.6	52.6	35.7	28.4	34.2	35.4
QSM (p⁢p⁢m)
18-40	0.0329	0.0609	0.0738	0.0717	0.1015	0.1150	0.0138	0.0130	0.0100	0.0279	0.0036	−0.0234	0.0241	0.0079	0.0119	0.0135	−0.0122
41-60	0.0400	0.0647	0.0713	0.0829	0.0984	0.1241	0.0134	0.0115	0.0025	0.0234	0.0085	−0.0226	0.0201	0.0079	0.0089	0.0099	−0.0110
61-80	0.0411	0.0705	0.0610	0.0738	0.0925	0.1249	0.0064	0.0089	−0.0034	0.0236	0.0061	−0.0243	0.0177	0.0100	0.0039	0.0096	−0.0091

Discussion

Our goal with the MASSP algorithm was to provide a fully automated method to delineate as many subcortical structures as possible on high-resolution structural MRI now available on 7T scanners. We modeled 17 distinct structures, taking into account location, shape, volume, and quantitative MRI contrasts to provide individual subject parcellations. Based on our results, we can be confident that the automated parcellation technique performs comparably to human experts, providing delineations within one or two voxels of the structure boundaries (dilated Dice overlap over 75% for all structures, including in aging groups). Results were nearly indistinguishable from expert delineations for eight major structures (Str, Tha, 4V, GPe, SN, RN, VTA, ic), and smaller structures retain high levels of overlap, comparable to trained human raters. This parcellation includes the most commonly defined structures (Str, Tha, SN, RN, STN) with overlap scores comparable to those previously reported (Garzón et al., 2018; Visser et al., 2016a; Eugenio Iglesias et al., 2013; Chakravarty et al., 2013; Patenaude et al., 2011). More importantly, it also includes structures seldom or never before considered in MRI atlases and parcellation methods, such as GPe, GPi, VTA, 3V, 4V, ic, fx, PAG, PPN, Cl. The technique handles structures of varying sizes well, as indicated by dilated overlap and boundary distance. Additional structures can be added, if they can be reliably delineated by expert raters on single-subject MRI at achievable resolutions. Some enhancement techniques such as building a multi-subject template (Pauli et al., 2018) or adding a denoising step (Bazin et al., 2019) may be beneficial. Co-registration to a high-precision atlas as in Ewert et al., 2018 may also improve the initial alignment over the MASSP group average template.

Age biases are present both in expert manual delineations and automated parcellation techniques. Age trajectories in volume and quantitative MR parameters indicate systematic shifts in contrast intensities and an increasing variability with age, associated with changing myelination, iron deposition, and brain atrophy (Draganski et al., 2011; Daugherty and Raz, 2013; Fjell et al., 2013; Keuken et al., 2017). These changes seem only to impact the parcellation accuracy for age groups beyond 60 years and age-matched priors did not provide specific improvements, thus indicating that an explicit modeling of age effects may be required to further improve parcellation quality in elderly populations. These results also point to exercising caution when applying automated parcellation methods to study morphometry in elderly or diseased populations, where measured differences may include biases. They also point out that while global volume and local thickness are indeed affected by such biases, quantitative MRI measures are much more robust. Note that this bias is likely present is many automated methods, although they have not been systematically investigated due to the extensive manual labor required. Interestingly, biases also exist in expert delineations: when the size or shape of a structure is refined in neuroanatomical studies, experts may become more or less conservative in their delineations. Automated methods provide a more objective measure in such case, as the source of their bias is explicitly encoded in the atlas prior delineations and computational model. Important applications of subcortical parcellation also include deep-brain stimulation surgery (Ewert et al., 2018), where the number of structures parcellated by MASSP can help neurosurgeons orient themselves more easily, although precise targeting will still require manual refinements, especially in neurodegenerative diseases.

We observed that dilated overlap, that is, the overlap of structures up to one voxel, provided a measure of accuracy largely independent of size, for automated or manual delineations. Imprecision in the range of one voxel in the boundary is to be expected due to partial voluming which impacts Dice overlap. The dilated overlap measure is a better representative of performance and indicates that conservative or inclusive versions of the subcortical regions can be obtained by eroding or dilating the estimated boundary by a single voxel. Such masks may be useful when separating functional MRI signals between neighboring nuclei or when locating smaller features inside a structure. Additionally, the Bayesian estimation framework provides voxel-wise probability values, which can also be used to further weight the contribution of each voxel within a region in subsequent analyses.

In summary, our method provides fast and accurate parcellation for subcortical structures of varying size, taking advantage of the high resolution offered by 7T and the specificity of quantitative MRI. The algorithm is based on an explicit model of structures given in a Bayesian framework and is free of tuning parameters. Given a different set of regions of interest or different populations, new priors can be automatically generated and used as the basis for the algorithm. If more MRI contrasts are available, the method can also be augmented to take them into account. The main requirement for the technique is a set of manual delineations of all the structures of interest in a small group of representative subjects. Performance may further improve with the number of included structures, as the number of distinct interfaces increases, refining in particular the intensity priors. In future works, we plan to include more structures or sub-structures and model the effects of age on the priors. We hope that the method, available in open source, will help neuroscience researchers to include more subcortical regions in their structural and functional imaging studies.

Materials and methodsData acquisition

Our parcellation method has been developed for the MP2RAGEME sequence (Caan et al., 2019). Briefly, the MP2RAGEME consists of two interleaved MPRAGEs with different inversions and four echoes in the second inversion. Based on these images, one can estimate quantitative MR parameters of R1, R2* and QSM. In this work, we used the following sequence parameters: inversion times TI1,2 = 670 ms, 3675.4 ms; echo times TE1 = 3 ms, TE2,1–4 = 3, 11.5, 19, 28.5 ms; flip angles FA1,2 = 4°, 4°; TRGRE1,2 = 6.2 ms, 31 ms; bandwidth = 404.9 MHz; TRMP2RAGE = 6778 ms; SENSE acceleration factor = 2; FOV = 205×205 x 164 mm; acquired voxel size = 0.70×0.7 x 0.7 mm; acquisition matrix was 292 × 290; reconstructed voxel size = 0.64×0.64 x 0.7 mm; turbo factor (TFE) = 150 resulting in 176 shots; total acquisition time = 19.53 min.

T1-maps were computed using a look-up table (Marques et al., 2010). T2*-maps were computed by least-squares fitting of the exponential signal decay over the multi-echo images of the second inversion. R1 and R2* maps were obtained as the inverse of T1 and T2*. For QSM, phase maps were pre-processed using iHARPERELLA (integrated phase unwrapping and background phase removal using the Laplacian) of which the QSM images were computed using LSQR (Li et al., 2014). Skull information was removed through creation of a binary mask using FSL’s brain extraction tool on the reconstructed uniform T1-weighted image and then applied to the quantitative contrasts (Smith, 2002). As all images were acquired as part of a single sequence, no co-registration of the quantitative maps was required (see Figure 9).

Figure 9.MP2RAGEME maps and delineations: quantitative R1 (left), quantitative R2* (middle), QSM (right).

Manual delineations for the 17 structures of interest are overlaid on all images.

Anatomical structure delineations

Manual delineations of subcortical structures were performed by two raters trained by an expert anatomist, according to protocols optimized to use the better contrast or combination of contrasts for each structure and to ensure a consistent approach across raters. The following 17 structures were defined on a group of 10 subjects (average age 24.4, eight female): striatum (Str), thalamus (Tha), lateral, 3rd and 4th ventricles (LV, 3V, 4V), amygdala (Amg), globus pallidus internal segment (GPi) and external segment (GPe), SN, STN, red nucleus (RN), ventral tegmental area (VTA), fornix (fx), internal capsule (ic), periaqueductal gray (PAG), pedunculopontine nucleus (PPN), and claustrum (Cl). Separate masks for left and right hemisphere were delineated except for 3V, 4V, and fx. In the following the algorithm treats each side separately, resulting in a total of 31 distinct structures (see Figure 1).

Anatomical interface priors

In order to inform the algorithm, we built a series of priors derived from the manual delineations. Each subject was first co-registered to a MP2RAGEME anatomical template built from 105 subjects co-aligned with the MNI2009b atlas (Fonov et al., 2011) with the SyN algorithm of ANTs (Avants et al., 2008) using successively rigid, affine, and non-linear transformations, high levels of regularization as recommended for the subcortex (Ewert et al., 2019) and mutual information as cost function.

The first computed prior is a prior of anatomical interfaces, recording the most likely location of boundaries between the different structures, defined as follows. Given two delineated structures i,j, let φi,φj be the signed distance functions to their respective boundary, that is, φi⁢(x) is the Euclidean distance of any given voxel to the boundary of i, with a negative sign inside the structure. Then we define the interface Bi|j with the distance function di|j:(1)di|j⁢(x)=min⁡(φi⁢(x),φj⁢(x)-δ,0)where δ is a scale parameter for the thickness of the interface. These interfaces functions are not symmetrical, as the intensity inside i next to j is generally different from the intensity inside j next to i. Based on this definition, the prior for a given interface based on N manual delineations is given by:(2)P(x∈Bi|j)∼12⁢π⁢σi|j2⁢(x)exp-12μi|j2⁢(x)σi|j2⁢(x)μi|j⁢(x)=1N⁢∑n∈Ndi|j,n⁢(x),σi|j⁢(x)=1N⁢∑n∈N(di|j,n⁢(x)-μi|j⁢(x))2+δ

These probability functions are calculated for all possible configurations including i|i, which represent the inside of each structure. We thus have a total of N2 functions, but only a few are non-zero at a given voxel x, and we may keep only the 16 largest values to account for any number of interfaces in 3D (Bazin et al., 2007). Finally, we need to scale the prior to be globally consistent with the priors below by assuming that the 95th percentile of the highest kept P(x∈Bi|j) values have a probability of 0.95. The scale parameter δ is set to one voxel, representing the expected amount of partial voluming. The resulting interface prior is shown in Figure 10A.

Figure 10.Anatomical interface (<bold>A</bold>) and skeleton (<bold>B</bold>) priors derived from the 10 manually delineated subjects.Anatomical skeleton priors

Next, we defined priors for the skeleton of each structure, representing their essential shape regardless of exact boundaries (Blum, 1973). As we are mostly interested in the most likely components of the skeleton or medial axis Si, we follow a simple method to estimate its location:(3)Si={x,|∇φi(x)|<12}

We define as si⁢(x) the signed distance function of this discrete skeleton, and define prior probabilities as above:(4)P(x∈Si)∼12⁢π⁢σi2⁢(x)exp-12μi2⁢(x)σi2⁢(x)μi⁢(x)=1N⁢∑n∈Nsi,n⁢(x)⁢σi⁢(x)=1N⁢∑n∈N(si,n⁢(x)-μi⁢(x))2+δ

The skeletons are defined inside each structure, which implies P(x∈Si)≤P(x∈Bi|i). To respect this relationship, we scale P(x∈Si) with the same factor as P(x∈Bi|i) but use P(x∈Si) when combining probabilities during the estimation stage. The obtained anatomical skeleton priors are given on Figure 10B.

Interface intensity priors

While anatomical priors already provide rich information, they are largely independent of the underlying MRI. From the co-aligned quantitative MRI maps and manual delineations, we defined intensity priors for every interface i|j, in the form of intensity histograms to ensure a flexible representation of intensity distributions. Given a quantitative contrast Rn⁢(x), we built a histogram Hi|j,n for each subject n and interface i|j. Histograms have 200 bins covering the entire intensity range within a radius of 10 mm from any of the delineated structures. To obtain an average histogram, we combine each histogram with a weighting function wn⁢(x) giving the likelihood of the subject’s intensity measurement compared to the group:(5)wi|j,n(x)=P(x∈Bi|j)12⁢π⁢σR2exp-12(Rn⁢(x)-μR⁢(x))2σR⁢(x)2where μR⁢(x) is the median of the Rn⁢(x) values at x, and σR⁢(x) is 1.349 times the inter-quartile range of Rn⁢(x). These are robust estimators of the mean and standard deviation, used here to avoid biases by intensity outliers. To further combine the R1, R2*, and QSM contrasts we take the geometric mean of the histogram probabilities: Hi|j⁢(x)=∏RHi|j⁢(R⁢(x))1/3.

Global volume priors

The last type of priors extracted from manual delineations are volume priors for each of the structure. Here, we assume a log-normal distribution for the volumes Vi and simply estimate the mean μV,i and standard deviation σV,i of log⁡Vi,n over the subjects.

Voxel-wise posterior probabilities

When parcellating a new subject, we first co-register its R1, R2*, and QSM maps jointly to the template and use the inverse transformation to deform the anatomical priors into subject space. Then we derive voxel-wise posteriors as follows:(6)P(x∈Bi|j|R(x))∼P(x∈Bi|j)Hi|j(x) if i≠jandP(x∈Bi|i|Si(x),R(x))∼max(P(x∈Bi|i),P(x∈Si)1/2)Hi|i(x)

Once again we should compute all possible combinations, but due to the multiplication of the priors we can restrict ourselves to the 16 highest probabilities previously estimated. To balance the contribution of the anatomical priors and the intensity histograms, we also need to normalize the intensity priors sampled on the subject’s intensities. We use the same approach, namely assuming that the 95th percentile of the highest kept Hi|j⁢(x) values have a probability of 0.95, separately for each contrast. The voxel-wise parcellation and posteriors obtained are shown in Figure 11A.

Figure 11.Successive parcellation results: (<bold>A</bold>) voxel-wise posteriors and parcellation, (<bold>B</bold>) diffused posteriors and parcellation, (<bold>C</bold>) topology-corrected posteriors and final region-growing parcellation.Markovian diffusion

The voxel-wise posteriors are independent from each other and do not reflect the continuous nature of the structures. The next step is to combine information from neighboring voxels. We define a sparse Markov Random Field model for the posteriors:(7)P(x∈Bi|j|R,S,C)=∑y∈C⁢(x)P(y∼x|R)P(y∈Bi|j|R,S,C)with P(y∼x|R)=∏Rexp-(R(y)-R(x))2/2σR2, where σR is the median of the standard deviations σi|j,R of the contrast histograms Hi|j⁢(R⁢(x)). The neighborhood C⁢(x) is defined as x itself and the four 26-connected neighboring voxels with highest probability P(y∼x|R), thus representing the neighbors most likely to be connected to x. The model is similar to a diffusion process and can be estimated with an iterated conditional modes (ICM) approach, updating sequentially the probabilities (Bazin and Pham, 2007):(8)P(x∈Bi|j|R,S,C)←∑y∈C⁢(x)P(y∼x|R)P(y∈Bi|j|R,S,C)from the initial voxel-wise posteriors until the ratio of changed parcellation labels decreases below 0.1, typically within 50–80 iterations. The diffused probabilities and parcellation are shown in Figure 11B.

Topology correction

The final step of the parcellation algorithm takes a global view of the individual structures, growing from the highest posterior values inside toward the boundaries. This region growing approach makes the implicit assumption that posterior maps should be monotonically decreasing from inside to outside, which is not necessarily the case. Therefore, we perform first a topology correction step on the individual structure posteriors P(x∈i|R,B,S,C)=maxi|jP(x∈Bi|j|R,S,C) with a fast marching algorithm (Bazin and Pham, 2007). While the corrected posterior is very similar to the original one (see Figure 11C), it ensures that all regions obtained by growing to a threshold have spherical object topology.

Anatomical region growing

Last, we turn the posteriors into optimized parcellations, by growing them concurrently (to avoid overlaps) until the target volume for each structure is reached. Given the volume Vi⁢(R,B,S,C) of the parcellation of the diffused and topology-corrected posteriors, we define the following target volume:(9)V^i=P⁢(Vi|μV,i,σV,i)⁢Vi+(1-P⁢(Vi|μV,i,σV,i))⁢exp⁡μV,itaking a weighted average of the volume estimated from the data and the prior volume. This approach ensures that even in extreme cases where some structures have low posteriors, they are still able to grow to a plausible size. The region growing algorithm is driven from the most likely voxels, defined as P(x∈i|R,B,S,C)-maxj≠iP(x∈j|R,B,S,C), and further modulated to follow isocontours of the skeleton prior:(10)P(x←y)∼P(y∈i|R,B,S,C)-maxj≠iP(y∈j|R,B,S,C)-|P(y∈Si)-P(x∈Si)|

Directionality of internal structures is a useful tool for understanding mechanical function in bones (Maquer et al., 2015). Here, we adapt this concept by using the skeleton isocontours as a representation of internal directionality, maintaining the intrinsic shape of structures. Thus, voxels with highest probability compared to the other structures and with similar distance to the internal skeleton are preferentially selected. The final parcellation is given in Figure 11C.

Validation metrics

To validate the method against manual expert delineations, we compared the MASSP results and the expert delineations with the following three measures:

The Dice overlap coefficient (Dice, 1945) D⁢(A,B)=2⁢A∩BA+B, which measures the strict overlap between voxels in both delineation;

The dilated overlap coefficient d⁢D⁢(A,B)=A∪d⁢(B)+B∪d⁢(A)A+B, where d(.) is a dilation of the delineation by one voxel, which measures the overlap between delineations allowing for one voxel of uncertainty;

The average surface distance a⁢s⁢d⁢(A,B), measuring the average distance between voxels on the surface boundary of the first delineation to the other one and reciprocally, which measures the distance between both delineations.

We computed all three measures for the manual delineations from the two independent raters, as well as the ratio of overlaps (automated over manual) and distances (manual over automated) to compare both performances, as detailed in the Results section.

Comparisons with other automated methods

To assess the performance of MASSP compared to existing parcellation tools, we ran Freesurfer (Fischl et al., 2002), FSL FIRST (Patenaude et al., 2011) and a multi-atlas registration approach (co-registering 9 of the 10 manually delineated subjects on the remaining one with ANTs [Avants et al., 2008] and labeling each structure by majority voting, similarly to the MAGeT Brain approach of Chakravarty et al., 2013). Freesurfer and FIRST were run on the skull-stripped R1 map, while the multi-atlas approach used all three R1, R2*, and QSM contrasts. All methods were compared in terms of Dice overlap, dilated overlap and average surface distance. We also assessed the presence of a systematic volume bias, defined as the average of the signed difference of the estimated structure volume to the manually delineated volume, normalized by the manually delineated volume.

Application to new MRI contrasts

Before applying MASSP to unseen contrasts, we need to convert its intensity prior histograms Hi|j,R to the new intensities. In order to perform this mapping, we first created a groupwise median of the HCP subjects, by co-registering every subject to the MASSP template using ANTs with non-linear registration and both T1w, T2w contrasts matched to the template’s R1 and R2* maps. The histogram bins are then updated as follows:(11)Hbin,i|j,R≡∑x|R⁢(x)∈binP(x∈Bi|j)Hi|j,R⁢1Hi|j,R⁢2⁣*Hi|j,Q⁢S⁢Madding the joint probability of the quantitative contrasts weighted by their importance for each interface to define the new intensity histograms. This model is essentially projecting the joint likelihood of the MASSP contrasts onto the new contrasts, assuming that the co-registration between the two is accurate enough. With these new histograms, we compared the test-retest reliability and overall agreement of MASSP with Freesurfer parcellations included in the HCP pre-processed data set.

Measurement of structure thickness

Finally, when comparing derived measures obtained over the lifespan with MASSP compared to manual delineations, we explored the utility of a shape thickness metric, based on the medial representation. Given the signed distance function φi of the structure boundary and s_i of the structure skeleton, the thickness is given by:(12)t⁢hi⁢(x)=2⁢(si⁢(x)-φi⁢(x))

Like in cortical morphometry, thickness is a local measure, defined everywhere inside the structure, and expected to provide additional information about anatomical variations. Indeed, a similar measure of shape thickness has recently been able to highlight subtle anatomical changes in depression (Ho et al., 2020).

Software implementation

The proposed method, Multi-contrast Anatomical Subcortical Structure Parcellation (MASSP), has been implemented as part of the Nighres toolbox (Huntenburg et al., 2018), using Python and Java for optimized processing. The software is available in open source from (release-1.3.0) and . A complete parcellation pipeline is included with the Nighres examples. Computations take under 30 min per subject on a modern workstation.

Acknowledgements

We thank Josephine Groot, Nikita Berendonk, Nicky Lute for their help collecting the AHEAD database, and Wietske van der Zwaag and Matthan Caan for their help in setting up the MP2RAGEME sequence. We also thank Steven Miletić and Dagmar Timmann for stimulating discussions around this topic, and all undergraduate students who contributed to the manual delineations. This work was supported by a NWO Vici grant (BF) and a NWO STW grant (AA, BF). HCP data were provided by the Human Connectome Project, WU-Minn Consortium (Principal Investigators: David Van Essen and Kamil Ugurbil; 1U54MH091657) funded by the 16 NIH Institutes and Centers that support the NIH Blueprint for Neuroscience Research; and by the McDonnell Center for Systems Neuroscience at Washington University.

Additional information

Competing interests

No competing interests declared

Author contributions

Conceptualization, Software, Formal analysis, Validation, Investigation, Visualization, Methodology, Writing - original draft

Data curation, Formal analysis, Validation, Investigation, Methodology, Writing - review and editing

Resources, Data curation, Software, Methodology, Writing - review and editing

Conceptualization, Methodology, Writing - review and editing

Conceptualization, Resources, Formal analysis, Supervision, Funding acquisition, Project administration, Writing - review and editing

Ethics

Human subjects: Informed consent and consent to publish, including consent to publish anonymized imaging data, was obtained for all subjects. Ethical approval was obtained with the University of Amsterdam Faculty of Social and Behavioral Sciences LAB Ethics Review Board, with ERB number 2016-DP-6897.

Additional files

Transparent reporting form

Data availability

The tool presented in this article is available in open source on Github (https://github.com/nighres/nighres). The atlases necessary to run the algorithm have been deposited on the University of Amsterdam FigShare (https://doi.org/10.21942/uva.12074175.v1 and https://doi.org/10.21942/uva.12301106.v2). A single sample subject data set has been deposited on the University of Amsterdam FigShare (https://doi.org/10.21942/uva.12280316.v2). All the measurements used to generate the figures included in the article have been deposited on the University of Amsterdam FigShare (https://doi.org/10.21942/uva.12452444.v1).

The following previously published datasets were used:

AlkemadeAMulderMJGrootJMIsaacsBRvan BerendonkNLuteNIsherwoodSJBazinP-LForstmannBU2020The Amsterdam Ultra-high field adult lifespan database (AHEAD): A freely available multimodal 7 Tesla submillimeter magnetic resonance imaging databaseFigShare / University of Amsterdam / Amsterdam University of Applied Sciences10.21942/uva.10007840.v1

Van EssenDCSmithSMBarchDMBehrensTEJYacoubEUgurbilK2013The WU-Minn Human Connectome ProjectWU-Minn HCP Retest Datahcp-young-adult

References

Ahsan

Allom

Gousias

Habib

Turkheimer

Free

Lemieux

Myers

Duncan

Brooks

Koepp

Hammers

2007

Volumes, spatial extents and a probabilistic atlas of the human basal ganglia and thalamus

NeuroImage38261270

10.1016/j.neuroimage.2007.06.004

17851093

Alkemade

Mulder

Groot

Isaacs

van Berendonk

Lute

Isherwood

Bazin

Forstmann

2020

The Amsterdam Ultra-high field adult lifespan database (AHEAD): A freely available multimodal 7 tesla submillimeter magnetic resonance imaging database

NeuroImage221

117200

10.1016/j.neuroimage.2020.117200

32745682

Andersen

Johnsen

Moos

2014

Iron deposits in the chronically inflamed central nervous system and contributes to neurodegeneration

Cellular and Molecular Life Sciences7116071622

10.1007/s00018-013-1509-8

24218010

Avants

Epstein

Grossman

Gee

2008

Symmetric diffeomorphic image registration with cross-correlation: evaluating automated labeling of elderly and neurodegenerative brain

Medical Image Analysis122641

10.1016/j.media.2007.06.004

17659998

Bazin

P-L

Ellingsen

Pham

2007

Digital Homeomorphisms in Deformable Registration

Karssemeijer

Lelieveldt

Information Processing in Medical ImagingBerlin, Heidelberg

Springer Berlin Heidelberg

211222

10.1007/978-3-540-73273-0_18

Bazin

P-L

Plessis

Fan

Villringer

Gauthier

2016

Vessel segmentation from quantitative susceptibility maps for local oxygenation venography

2016 IEEE 13th International Symposium on Biomedical Imaging (ISBI)

11351138

10.1109/ISBI.2016.7493466

Bazin

Alkemade

van der Zwaag

Caan

Mulder

Forstmann

2019

Denoising High-Field Multi-Dimensional MRI with local complex PCA

Frontiers in Neuroscience13

10.3389/fnins.2019.01066

31649500

Bazin

Pham

2007

Topology correction of segmented medical images using a fast marching algorithm

Computer Methods and Programs in Biomedicine88182190

10.1016/j.cmpb.2007.08.006

17942182

Bazin

Pham

2008

Homeomorphic brain image segmentation with topological and statistical atlases

Medical Image Analysis12616625

10.1016/j.media.2008.06.008

18640069

Blum

1973

Biological shape and visual science. I

Journal of Theoretical Biology38205287

10.1016/0022-5193(73)90175-6

4689997

Caan

MWA

Bazin

Marques

Hollander

Dumoulin

Zwaag

2019

MP2RAGEME: T 1 , T 2 * , and QSM mapping in one sequence at 7 tesla

Human Brain Mapping4017861798

10.1002/hbm.24490

30549128

Chakravarty

Bertrand

Hodge

Sadikot

Collins

2006

The creation of a brain atlas for image guided neurosurgery using serial histological data

NeuroImage30359376

10.1016/j.neuroimage.2005.09.041

16406816

Chakravarty

Steadman

van Eede

Calcott

Shaw

Raznahan

Collins

Lerch

2013

Performing label-fusion-based segmentation using multiple automatically generated templates: maget brain: label fusion segmentation using automatically generated templates

Human Brain Mapping3426352654

10.1002/hbm.22092

22611030

Daugherty

Raz

2013

Age-related differences in iron content of subcortical nuclei observed in vivo: a meta-analysis

NeuroImage70113121

10.1016/j.neuroimage.2012.12.040

23277110

de Hollander

Keuken

van der Zwaag

Forstmann

Trampel

2017

Comparing functional MRI protocols for small, iron-rich basal ganglia nuclei such as the subthalamic nucleus at 7 T and 3 T

Human Brain Mapping3832263248

10.1002/hbm.23586

28345164

Dice

1945

Measures of the amount of ecologic association between species

Ecology26297302

10.2307/1932409

Draganski

Ashburner

Hutton

Kherif

Frackowiak

Helms

Weiskopf

2011

Regional specificity of MRI contrast parameter changes in normal ageing revealed by voxel-based quantification (VBQ)

NeuroImage5514231434

10.1016/j.neuroimage.2011.01.052

21277375

Eugenio Iglesias

Rory Sabuncu

Van Leemput

2013

A unified framework for cross-modality multi-atlas segmentation of brain MRI

Medical Image Analysis1711811191

10.1016/j.media.2013.08.001

24001931

Ewert

Plettig

Chakravarty

Collins

Herrington

Kühn

Horn

2018

Toward defining deep brain stimulation targets in MNI space: a subcortical atlas based on multimodal MRI, histology and structural connectivity

NeuroImage170271282

10.1016/j.neuroimage.2017.05.015

28536045

Ewert

Horn

Finkel

Kühn

Herrington

2019

Optimization and comparative evaluation of nonlinear deformation algorithms for atlas-based segmentation of DBS target nuclei

NeuroImage184586598

10.1016/j.neuroimage.2018.09.061

30267856

Fischl

Salat

Busa

Albert

Dieterich

Haselgrove

van der Kouwe

Killiany

Kennedy

Klaveness

Montillo

Makris

Rosen

Dale

2002

Whole brain segmentation: automated labeling of neuroanatomical structures in the human brain

Neuron33341355

10.1016/s0896-6273(02)00569-x

11832223

Fjell

Westlye

Grydeland

Amlien

Espeseth

Reinvang

Raz

Holland

Dale

Walhovd

Alzheimer Disease Neuroimaging Initiative

2013

Critical ages in the life course of the adult brain: nonlinear subcortical aging

Neurobiology of Aging3422392247

10.1016/j.neurobiolaging.2013.04.006

23643484

Fonov

Evans

Botteron

Almli

McKinstry

Collins

Brain Development Cooperative Group

2011

Unbiased average age-appropriate atlases for pediatric studies

NeuroImage54313327

10.1016/j.neuroimage.2010.07.033

20656036

Forstmann

Keuken

Schafer

Bazin

Alkemade

Turner

2014

Multi-modal ultra-high resolution structural 7-Tesla MRI data repository

Scientific Data1

140050

10.1038/sdata.2014.50

25977801

Forstmann

de Hollander

van Maanen

Alkemade

Keuken

2016

Towards a mechanistic understanding of the human subcortex

Nature Reviews Neuroscience185765

10.1038/nrn.2016.163

27974841

Forstmann

Isaacs

Temel

2017

Ultra high field MRI-Guided deep brain stimulation

Trends in Biotechnology35904907

10.1016/j.tibtech.2017.06.010

28941469

Frazier

Chiu

Breeze

Makris

Lange

Kennedy

Herbert

Bent

Koneru

Dieterich

Hodge

Rauch

Grant

Cohen

Seidman

Caviness

Biederman

2005

Structural brain magnetic resonance imaging of limbic and thalamic volumes in pediatric bipolar disorder

American Journal of Psychiatry16212561265

10.1176/appi.ajp.162.7.1256

15994707

Garzón

Sitnikov

Bäckman

Kalpouzos

2018

Automated segmentation of midbrain structures with high iron content

NeuroImage170199209

10.1016/j.neuroimage.2017.06.016

28602813

Herting

Johnson

Mills

Vijayakumar

Dennison

Liu

Goddings

Dahl

Sowell

Whittle

Allen

Tamnes

2018

Development of subcortical volumes across adolescence in males and females: a multisample study of longitudinal changes

NeuroImage172194205

10.1016/j.neuroimage.2018.01.020

29353072

Hill

Grutzendler

2018

Lifelong cortical myelin plasticity and age-related degeneration in the live mammalian brain

Nature Neuroscience21683695

10.1038/s41593-018-0120-6

29556031

Gutman

Pozzi

Grabe

Hosten

Wittfeld

Völzke

Baune

Dannlowski

Förster

Grotegerd

Redlich

Jansen

Kircher

Krug

Meinert

Nenadic

Opel

Dinga

Veltman

Schnell

Veer

Walter

Gotlib

Sacchet

Aleman

Groenewold

Stein

Walter

Ching

CRK

Jahanshad

Ragothaman

Isaev

Zavaliangos‐Petropulu

Thompson

Sämann

Schmaal

2020

Subcortical shape alterations in major depressive disorder: findings from the ENIGMA major depressive disorder working group

Human Brain Mapping1

e24988

10.1002/hbm.24988

Huntenburg

Steele

Bazin

2018

Nighres: processing tools for high-resolution neuroimaging

GigaScience7

giy082

10.1093/gigascience/giy082

29982501

Iglesias

Insausti

Lerma-Usabiaga

Bocchetta

Van Leemput

Greve

van der Kouwe

Fischl

Caballero-Gaudes

Paz-Alonso

Alzheimer's Disease Neuroimaging Initiative

2018

A probabilistic atlas of the human thalamic nuclei combining ex vivo MRI and histology

NeuroImage183314326

10.1016/j.neuroimage.2018.08.012

30121337

Spronk

Kulkarni

Repovš

Anticevic

Cole

2019

Mapping the human brain's cortical-subcortical functional network organization

NeuroImage1853557

10.1016/j.neuroimage.2018.10.006

30291974

Johansen-Berg

2013

Human connectomics - what will the future demand?

NeuroImage80541544

10.1016/j.neuroimage.2013.05.082

23727322

Keuken

Bazin

Schäfer

Neumann

Turner

Forstmann

2013

Ultra-high 7T MRI of structural age-related changes of the subthalamic nucleus

Journal of Neuroscience3348964900

10.1523/JNEUROSCI.3241-12.2013

23486960

Keuken

Bazin

Backhouse

Beekhuizen

Himmer

Kandola

Lafeber

Prochazkova

Trutti

Schäfer

Turner

Forstmann

2017

Effects of aging on T₁, T₂*, and QSM MRI values in the subcortex

Brain Structure & Function22224872505

10.1007/s00429-016-1352-4

28168364

Koshiyama

Fukunaga

Okada

Yamashita

Yamamori

Yasuda

Fujimoto

Ohi

Fujino

Watanabe

Kasai

Hashimoto

2018

Role of subcortical structures on cognitive and social function in schizophrenia

Scientific Reports8

1183

10.1038/s41598-017-18950-2

29352126

Avram

Xiao

Liu

2014

Integrated Laplacian-based phase unwrapping and background phase removal for quantitative susceptibility mapping

NMR in Biomedicine27219227

10.1002/nbm.3056

24357120

Makowski

Béland

Kostopoulos

Bhagwat

Devenyi

Malla

Joober

Lepage

Chakravarty

2018

Evaluating accuracy of striatal, pallidal, and thalamic segmentation methods: comparing automated approaches to manual delineation

NeuroImage170182198

10.1016/j.neuroimage.2017.02.069

28259781

Maquer

Musy

Wandel

Gross

Zysset

2015

Bone volume fraction and fabric anisotropy are better determinants of trabecular bone stiffness than other morphological variables

Journal of Bone and Mineral Research3010001008

10.1002/jbmr.2437

25529534

Marquand

Haak

Beckmann

2017

Functional corticostriatal connection topographies predict goal directed behaviour in humans

Nature Human Behaviour1

0146

10.1038/s41562-017-0146

28804783

Marques

Kober

Krueger

van der Zwaag

Van de Moortele

Gruetter

2010

MP2RAGE, a self bias-field corrected sequence for improved segmentation and T1-mapping at high field

NeuroImage4912711281

10.1016/j.neuroimage.2009.10.002

19819338

McCoy

Wakefield

Akey

2017

Impacts of Neanderthal-Introgressed sequences on the landscape of human gene expression

Cell168916927

10.1016/j.cell.2017.01.038

28235201

Miletić

Bazin

Weiskopf

van der Zwaag

Forstmann

Trampel

2020

fMRI protocol optimization for simultaneously studying small subcortical and cortical Areas at 7T

NeuroImage219

116992

10.1016/j.neuroimage.2020.116992

32480037

Mosley

Smith

Coyne

Silburn

Breakspear

Perry

2018

The site of stimulation moderates neuropsychiatric symptoms after subthalamic deep brain stimulation for Parkinson's disease

NeuroImage: Clinical189961006

10.1016/j.nicl.2018.03.009

29876284

Patenaude

Smith

Kennedy

Jenkinson

2011

A Bayesian model of shape and appearance for subcortical brain segmentation

NeuroImage56907922

10.1016/j.neuroimage.2011.02.046

21352927

Pauli

Nili

Tyszka

2018

A high-resolution probabilistic in vivo atlas of human subcortical brain nuclei

Scientific Data5

180063

10.1038/sdata.2018.63

29664465

Qiu

Brown

Fischl

Miller

2010

Atlas generation for subcortical and ventricular structures with its applications in shape analysis

IEEE Transactions on Image Processing1915391547

10.1109/TIP.2010.2042099

20129863

Sitek

Gulban

Calabrese

Johnson

Lage-Castellanos

Moerel

Ghosh

De Martino

2019

Mapping the human subcortical auditory system using histology. post mortem MRI and in vivo MRI at 7T

bioRxiv10.1101/568139

Smith

2002

Fast robust automated brain extraction

Human Brain Mapping17143155

10.1002/hbm.10062

12391568

Steiner

Tseng

2017

Handbook of Basal Ganglia Structure and Function

Steiner

Tseng

Handbook of Behavioral Neuroscience24

Elsevier

11036

Tardif

Gauthier

Steele

Bazin

Schäfer

Schaefer

Turner

Villringer

2016

Advanced MRI techniques to improve our understanding of experience-induced neuroplasticity

NeuroImage1315572

10.1016/j.neuroimage.2015.08.047

26318050

Turner

2019

Myelin and modeling: bootstrapping cortical microcircuits

Frontiers in Neural Circuits13

10.3389/fncir.2019.00034

31133821

Van Essen

Smith

Barch

Behrens

Yacoub

Ugurbil

WU-Minn HCP Consortium

2013

The WU-Minn human connectome project: an overview

NeuroImage806279

10.1016/j.neuroimage.2013.05.041

23684880

Visser

Keuken

Douaud

Gaura

Bachoud-Levi

Remy

Forstmann

Jenkinson

2016a

Automatic segmentation of the striatum and globus pallidus using MIST: multimodal image segmentation tool

NeuroImage125479497

10.1016/j.neuroimage.2015.10.013

26477650

Visser

Keuken

Forstmann

Jenkinson

2016b

Automated segmentation of the Substantia Nigra, subthalamic nucleus and red nucleus in 7T data at young and old age

NeuroImage139324336

10.1016/j.neuroimage.2016.06.039

27349329

Wang

Poirier

Guo

Parrent

Peters

Khan

Styner

Angelini

2016

Generation and evaluation of an ultra-high-field atlas with applications in DBS planning

Proceedings of SPIE Medical Imaging

Weiskopf

Suckling

Williams

Correia

Inkster

Tait

Ooi

Bullmore

Lutti

2013

Quantitative multi-parameter mapping of R1, PD(*), MT, and R2(*) at 3T: a multi-center validation

Frontiers in Neuroscience7

10.3389/fnins.2013.00095

23772204

Xiao

Fonov

Bériault

Al Subaie

Chakravarty

Sadikot

Pike

Collins

2015

Multi-contrast unbiased MRI atlas of a parkinson's disease population

International Journal of Computer Assisted Radiology and Surgery10329341

10.1007/s11548-014-1068-y

24841147

Yeatman

Wandell

Mezer

2014

Lifespan maturation and degeneration of human brain white matter

Nature Communications5

4932

10.1038/ncomms5932

25230200

Yelnik

Bardinet

Dormont

Malandain

Ourselin

Tandé

Karachi

Ayache

Cornu

Agid

2007

A three-dimensional, histological and deformable atlas of the human basal ganglia. I. Atlas construction based on immunohistochemical and MRI data

NeuroImage34618638

10.1016/j.neuroimage.2006.09.026

17110133

Zhang

Wei

Cronin

Yan

Liu

2018

Longitudinal atlas for normative human brain development and aging over the lifespan using quantitative susceptibility mapping

NeuroImage171176189

10.1016/j.neuroimage.2018.01.008

29325780

10.7554/eLife.59430.sa1

Decision letter

Verstynen

Timothy

Reviewing EditorCarnegie Mellon UniversityUnited States

Verstynen

Timothy

ReviewerCarnegie Mellon UniversityUnited StatesNeumann

Wolf-Julian

ReviewerCharité - Universitätsmedizin BerlinGermany

In the interests of transparency, eLife publishes the most substantive revision requests and the accompanying author responses.

Acceptance summary:

There was unanimous agreement that this method has strong potential to be a new "workhorse" tool in human neuroimaging that could substantially advance our ability to measure brain structures that are largely overlooked due to problems with segmentation.

Decision letter after peer review:

Thank you for submitting your article "Multi-contrast Anatomical Subcortical Structures Parcellation" for consideration by eLife. Your article has been reviewed by three peer reviewers, including Timothy Verstynen as the Reviewing Editor and Reviewer #1, and the evaluation has been overseen by Michael Frank as the Senior Editor. The following individual involved in review of your submission has agreed to reveal their identity: Wolf-Julian Neumann (Reviewer #2).

The reviewers have discussed the reviews with one another and the Reviewing Editor has drafted this decision to help you prepare a revised submission.

We would like to draw your attention to changes in our revision policy that we have made in response to COVID-19 (https://elifesciences.org/articles/57162). Specifically, we are asking editors to accept without delay manuscripts, like yours, that they judge can stand as eLife papers without additional data, even if they feel that they would make the manuscript stronger. Thus the revisions requested below only address clarity and presentation.

Summary:

In this study, Bazin and colleagues propose a novel segmentation algorithm for parcelling subcortical regions of the human brain that was developed from multiple MRI measures derived from the M2RAGEME sequence acquired on a 7T MRI system. The key advancement of this approach is a reliable segmentation of more subcortical areas (17 regions) in native space than what is possible with currently available methods. The authors validate their algorithm by comparing against age-related measures.

This manuscript was reviewed by three experts in the field, who found that this method has strong potential to be a new "workhorse" tool in human neuroimaging that could substantially advance our ability to measure brain structures that are largely overlooked due to problems with segmentation. The main criticisms of the work are largely centered on how the method is evaluated and implemented, rather than fundamental concerns with the validity of the method itself.

Essential revisions:

1) Benchmarks.

All three reviewers had concerns about the nature of the tests for the new method.

Reviewer 1 was particularly concerned that, while a critical advancement of this method is the ability to segment many more regions than previous subcortical atlases, there are still many regions that overlap with existing segmentation tools. Knowing how the reliability of this new approach compares to previous automatic segmentation methods is crucial in being able to know how to trust the overall reliability of the method. The authors should make a direct benchmark against previous methods where they have overlap.

Reviewer 2 thought that the work would certainly benefit from an additional step of out-of-center / out-of-cohort validation analysis. Though they had no serious concern that performance would be unsatisfactory, it would still highlight the extensibility of the method.

Reviewer 3 shared a similar concern, pointing out that automatized methods are usually sensitive to the number of subjects used to build the parcellation, with results from a bigger training cohort being potentially more robust and generalizable. One of the strongest points of the automated method presented in this paper is the adoption of a Bayesian approach, which usually works efficiently for small sample sizes and allows to update previous results when new data comes. Still, it could be highly illustrative to show the performance of the current method depending on the initial training size. From the same set of delineations of the 105 subjects used to test the age bias, what if the authors show the predicted performance from generating the priors on a training set varying its size?

2) Aging analysis.

All three reviewers were confused as to the purpose for and implementation of the aging analysis included in the paper.

Reviewer 1 said that the analysis of the aging effects on the segmentations seemed oddly out of place. It wasn't clear if this is being used to vet the effectiveness of the algorithm (i.e., its ability to pick up on patterns of age-related changes) or the limitations of the algorithm (i.e., the segmentation effectiveness decreases in populations with lower across-voxel contrast). What exactly is the goal with this analysis? Also, why is it limited to only a subset of the regions output from the algorithm?

Reviewer 2 thought that the most important limitation, as acknowledged by the authors, is the bias from anatomical variation through age or disease. The algorithm is shown to be affected by age and most certainly will be affected by contrast and size changes in neurodegenerative disorders. Broader benchmark tests, as proposed above, would likely address this concern.

Reviewer 3 pointed out that, from Figure 4, it is clear how estimated Dice coefficients decrease with age. As it is well noted by the authors, this is likely caused due to the fact that the priors were built from 10 subjects that had an average age of 24.4 years and thus, the highest predicted performance rates are reflected for subjects whose age range (18-40) lies around this average prior age. The authors mentioned in the paper that they plan on modelling the effects of age in the priors in future work. However, they could already address this question in the current work. Since the data used to test this age bias has already been manually delineated, what if the authors generate new priors for this set of delineations, including subjects from all ages, and test whether the predicted Dice coefficients still depend on age, in the same way as was done in Figure 4?

3) Clarity of the algorithm.

Reviewers 1 and 3 had concerns about details of the algorithm itself.

Reviewer 1 thought that, because of the difficulty of the parcellation problem, the algorithm being used is quite complex. The authors do a good job showing the output of each stage of the process (Figure 7 and Figure 8), but it would substantially help general readers to have a schematic of the logic of the algorithm itself.

Reviewer 3 pointed out that it is unclear what is the value for the scale parameter δ that appears in the priors? Is that a free parameter? If so, do results change when this parameter varies? This seems to be a critical aspect of the process (at least insofar as the precision of the results).

[Editors' note: further revisions were suggested prior to acceptance, as described below.]

Thank you for resubmitting your article "Multi-contrast Anatomical Subcortical Structures Parcellation" for consideration by eLife. Your revised article has been reviewed by three peer reviewers, including Timothy Verstynen as the Reviewing Editor and Reviewer #1, and the evaluation has been overseen by Michael Frank as the Senior Editor. The following individual involved in review of your submission has agreed to reveal their identity: Wolf-Julian Neumann (Reviewer #2).

The reviewers have discussed the reviews with one another and the Reviewing Editor has drafted this decision to help you prepare a revised submission.

Summary:

The reviewers felt that the revised manuscript is much stronger and focused. There remains only a minor point raised by reviewer #2 that we ask you address.

Essential revisions:

Reviewer #2 (see below) would like to see a more elaborate comparison of the STN segmentation to previous recent methods, given that these sorts of subcortical parcellation methods are likely to be of interest to DBS researchers. While a comparison to the Ewert method (or similar) would be nice, a brief discussion of a subjective comparison of these results would be sufficient to address this concern.

Reviewer #1:

The authors have adequately addressed all of my concerns. Well done.

Reviewer #2:

This is a revised manuscript on a 7T based segmentation approach. My main concern in the primary submission was that at this point the complexity of the data acquisition in combination with the computational approach makes it relatively niche. Other concerns regarded additional validation runs and aging, which the authors have now provided. My feeling is that the authors have made an effort to address the major points raised in the first revision.

There is one minor point that still remains puzzling for me. The fact that the STN is delineated so poorly, when compared to other structures. Given its use as a target in hundreds of thousands of patients for deep brain stimulation, automatized STN segmentation has been validated. The authors cited Ewert et al., which combines a simple normalization with an atlas in MNI space and reached similar and better performance when compared to the presented results here. I would have liked to see the results from this paper reproduced as a comparison here, but if the authors decide not to, I would at least like to invite the authors to discuss why the STN is troublesome for the algorithm and how this could affect use for surgical planning in the future, when 7T becomes available in clinics.

Additionally, I found this a little strange: Figure 3 caption: compared to the most expert of the two human raters.

Reviewer #3:

The authors have addressed all the concerns that I had in the previous version of the manuscript. Important changes in this revision included a benchmark against existing automated parcellation tools, a validation analysis using a test-retest sample from the Human Connectome Project and a thorough examination of training sample size and age biases. All of these changes have significantly increased the quality of the paper and more importantly, provided more clarity and evidence for the benefits of the proposed algorithm for subcortical parcellation. As a consequence, I am more than pleased to recommend the current version of the manuscript for publication.

10.7554/eLife.59430.sa2

Author response

Essential revisions:

1) Benchmarks.

All three reviewers had concerns about the nature of the tests for the new method.

We agree with reviewer 1 that comparing to other previously published methods is useful, as few of them provide tables of validation scores. We added a comparison with three popular tools: a multi-atlas registration scheme building directly on our manual delineations, FSL First, and Freesurfer. The results are summarized in a new Table 2, and detailed further in the Materials and methods and Results sections.

In Results:

“Comparison to other automated methods

To provide a basis for comparison, we applied other freely available methods for subcortical structure parcellation to the same ten subjects. MASSP performs similarly or better than Freesurfer, FSL First and a multi-atlas registration using ANTs. Multi-atlas registration provides high accuracy in most structures as well, but is biased toward under-estimating the size of smaller and elongated structures where overlap is systematically reduced across the individual atlas subjects. Multi-atlas registration is also quite computationally intensive when using multiple contrasts at high resolution. Finally, MASSP provides many more structures than Freesurfer and FSL First, and can be easily applied to new structures based on additional manual delineations.”

In Materials and methods:

“Comparisons with other automated methods

To assess the performance of MASSP compared to existing parcellation tools, we ran Freesurfer (Fischl et al., 2002), FSL First (Patenaude et al., 2011) and a multi-atlas registration approach (coregistering 9 of the 10 manually delineated subjects on the remaining one with ANTs (Avants et al., 2008) and labelling each structure by majority voting, similarly to the MAGeT Brain approach of Chakravarty et. al (2013)). Freesurfer and First were run on the skull-stripped R1 map, while the multi-atlas approach used all three R1, R2* and QSM contrasts. All methods were compared in terms of Dice overlap, and we also assessed the presence of a systematic volume bias, defined as the average of the signed difference of the estimated structure volume to the manually delineated volume, normalized by the manually delineated volume.”

We thank reviewer 2 for this thought, which led us to define an appropriate methodology to expand the algorithm to other contrasts. While MASSP is based on quantitative MRI, it is not entirely straightforward to simulate the contrast of various MR sequences based on these, as other sources of contrasts and artifacts often influence the imaging. Instead, we took a data-driven approach, coregistering templates with the contrasts of interest and building a statistical mapping of intensities for each defined interface in the MASSP atlas. With this approach, we were able to successfully parcellate data from the Human Connectome Project. We made the following additions to the Materials and methods and Results sections:

In Results:

“Application to new MRI contrasts

Quantitative MRI has only become recently applicable in larger studies, thanks in part to the development of integrated multi-parameter sequences (Weiskopf et al., 2013; Caan et al., 2019). Many data sets, including large-scale open databases, use more common T1- and T2-weighted MRI. In order to test the applicability of MASSP to such contrasts, we obtained the test-retest subset of the Human Connectome Project (HCP, Van Essen et al., 2013) and applied MASSP to the 45 preprocessed and skull-stripped T1- and T2-weighted images from each of the two test and retest sessions. While performing manual delineations on the new contrasts would be preferable, the model is already rich enough to provide stable parcellations. Test-retest reproducibility is similarly high for MASSP and Freesurfer, and are generally in agreement, see Figure 5 and Table 3.”

In Materials and methods:

“Application to new MRI contrasts

We thank Reviewer 3 for this suggestion. We used the delineations of the five subcortical structures defined on the entire cohort to define atlases of increasing size, from 3 to 18 subjects. Prior work from us and others (e.g. Bazin et al., 2008; Iglesias et al., 2013) have shown that Bayesian approaches are generally very efficient with regard to the size of the training cohort, and we found indeed that performance stabilized for all structures at 8 subjects. We added the experiment to the manuscript as follows in Results:

“Biases due to atlas size

[…] First, we investigated the impact of atlas size. We randomly assigned half of the subjects from each decade to two groups, and built atlas priors from subsets of 3, 5, 8, 10, 12, 15, and 18 subjects from the first group. The subjects used in the atlas were taken randomly from each decade (18-30, 31-40, 41-50, 51-60, 61-70, 71-80), so as to maximize the age range represented in each atlas. Atlases of increasing size were constructed by adding subjects to previous atlases, so that atlases of increasing complexity include all subjects from simpler atlases. Results applying these atlases to parcellate the second group are given in Figure 6. As in previous studies (Iglesias et al., 2013; Bazin et al., 2008), performance quickly stabilized with atlases of more than five subjects (no significant difference in Welch's t-tests between using 18 subjects or any subset of 8 or more for all structures and measures).”

2) Aging analysis.

All three reviewers were confused as to the purpose for and implementation of the aging analysis included in the paper.

We thank the reviewers for their detailed comments and suggestions. Indeed, we realize that the aging analysis was not clearly motivated, and that the experiment of Figure 4 was not very informative. Our goal here was to assess the ability of the algorithm to parcellate data from older subjects, a step seldom taken when validating algorithms. In this revision we replaced the experiment of Figure 4 by a more thorough test of biases: using only the five structures for which we have a complete set of delineations, we tested systematically the impact of atlas age range versus testing age group. The results, reproduced below, indicate that while there is a decrease in performance in subjects above 60 year old, the decrease appears largely independent of the choice of the atlas age group. Thus we conclude that the method accuracy does decrease in subjects above 60 year old and likely in patients with neurodegenerative disease. This effect is not merely a reflection of bias in the atlas, but rather the expression of increasing variability in brain MRI. Note however that the following experiment on estimating morphometry and quantitative MRI from automated versus manual parcellations shows a good stability of local measures across age groups, even if structure boundaries and volumes are not perfectly estimated.

In Results:

“Biases due to age differences

To more specifically test the influence of age on parcellation accuracy, we defined six age groups by decade and randomly selected 10 subjects from each group. Each set of subjects was used as priors for the five structures above, and applied to the other age groups. Results are summarized in Figure 7. Examining this age bias, we can see a decrease in performance when parcellating subjects in the range of 60 to 80 years of age. The choice of priors seems to have a limited impact, which varies across structures. Interestingly, using priors from a similar age group is not particularly beneficial.”

3) Clarity of the algorithm.

Reviewers 1 and 3 had concerns about details of the algorithm itself.

We thank reviewer 1 for this valuable suggestion. We added the following explanatory diagram at the beginning of Results:

We thank the reviewer for noticing this omission: δ represents the amount of partial voluming at the interface. In theory it could be tuned (in particular to be increased at smoother boundaries such as the thalamus-internal capsule interface), but setting it to the imaging scale provided the most consistent results across structures. We added this mention to the Materials and methods:

The scale parameter is set to 1 voxel, representing the expected amount of partial voluming.

[Editors' note: further revisions were suggested prior to acceptance, as described below.]

Essential revisions:

In the article by Ewert et al., the parcellation is performed by co-registration to a very precise high resolution atlas. In their experiment (see their Table 2), they report average Cohen's Kappa of 0.4467 and 0.5240 for the STN and the GPi respectively. We recomputed the corresponding metric for our leave-one-out experiment and obtained 0.4843 and 0.5452 respectively, which is within the same range of accuracy. These structures, the STN particularly, are notoriously difficult to parcellate due to low T1 contrast with the WM of the internal capsule and the proximity of nuclei with similar T2 and T2* contrast (SN, RN GPe). However, the relatively high performance of the atlas co-registration by Ewert et al., 2018 and of multi-atlas co-registration for these structures indicates that further performance improvements may be gained by using a more anatomically precise template. We added the following to the discussion:

“Some enhancement techniques such as building a multi-subject template (Pauli et al., 2018) or adding a denoising step (Bazin et al., 2019) may be beneficial. Co-registration to a high-precision atlas as in (Ewert et al., 2018) may also improve the initial alignment over the MASSP group average template.”

and later on, when discussing applications:

“Important applications of subcortical parcellation also include deep-brain stimulation surgery (Ewert et al., 2018), where the number of structures parcellated by MASSP can help neurosurgeons orient themselves more easily, although precise targeting will still require manual refinements, especially in neurodegenerative diseases.”

Reviewer #2:

[…]

Additionally, I found this a little strange: Figure 3 caption: compared to the most expert of the two human raters.

We changed the text to "compared to the human rater with most neuroanatomical expertise." Here the goal was to compare to our most skilled manual delineations, rather than a consensus between expert and trainee.