EASI-FISH for thick tissue defines lateral hypothalamus spatio-molecular organization.

Determining the spatial organization and morphological characteristics of molecularly defined cell types is a major bottleneck for characterizing the architecture underpinning brain function. We developed Expansion-Assisted Iterative Fluorescence In Situ Hybridization (EASI-FISH) to survey gene expression in brain tissue, as well as a turnkey computational pipeline to rapidly process large EASI-FISH image datasets. EASI-FISH was optimized for thick brain sections (300 µm) to facilitate reconstruction of spatio-molecular domains that generalize across brains. Using the EASI-FISH pipeline, we investigated the spatial distribution of dozens of molecularly defined cell types in the lateral hypothalamic area (LHA), a brain region with poorly defined anatomical organization. Mapping cell types in the LHA revealed nine spatially and molecularly defined subregions. EASI-FISH also facilitates iterative reanalysis of scRNA-seq datasets to determine marker-genes that further dissociated spatial and morphological heterogeneity. The EASI-FISH pipeline democratizes mapping molecularly defined cell types, enabling discoveries about brain organization.

Multimodal mapping of cell types and projections in the central nucleus of the amygdala.

The central nucleus of the amygdala (CEA) is a brain region that integrates external and internal sensory information and executes innate and adaptive behaviors through distinct output pathways. Despite its complex functions, the diversity of molecularly defined neuronal types in the CEA and their contributions to major axonal projection targets have not been examined systematically. Here, we performed single-cell RNA-sequencing (scRNA-seq) to classify molecularly defined cell types in the CEA and identified marker genes to map the location of these neuronal types using expansion-assisted iterative fluorescence in situ hybridization (EASI-FISH). We developed new methods to integrate EASI-FISH with 5-plex retrograde axonal labeling to determine the spatial, morphological, and connectivity properties of ~30,000 molecularly defined CEA neurons. Our study revealed spatiomolecular organization of the CEA, with medial and lateral CEA associated with distinct molecularly defined cell families. We also found a long-range axon projection network from the CEA, where target regions receive inputs from multiple molecularly defined cell types. Axon collateralization was found primarily among projections to hindbrain targets, which are distinct from forebrain projections. This resource reports marker gene combinations for molecularly defined cell types and axon-projection types, which will be useful for selective interrogation of these neuronal populations to study their contributions to the diverse functions of the CEA.

Fast predictive simple geodesic regression.

Deformable image registration and regression are important tasks in medical image analysis. However, they are computationally expensive, especially when analyzing large-scale datasets that contain thousands of images. Hence, cluster computing is typically used, making the approaches dependent on such computational infrastructure. Even larger computational resources are required as study sizes increase. This limits the use of deformable image registration and regression for clinical applications and as component algorithms for other image analysis approaches. We therefore propose using a fast predictive approach to perform image registrations. In particular, we employ these fast registration predictions to approximate a simplified geodesic regression model to capture longitudinal brain changes. The resulting method is orders of magnitude faster than the standard optimization-based regression model and hence facilitates large-scale analysis on a single graphics processing unit (GPU). We evaluate our results on 3D brain magnetic resonance images (MRI) from the ADNI datasets.

Joint Intensity Fusion Image Synthesis Applied to Multiple Sclerosis Lesion Segmentation.

We propose a new approach to Multiple Sclerosis lesion segmentation that utilizes synthesized images. A new method of image synthesis is considered: joint intensity fusion (JIF). JIF synthesizes an image from a library of deformably registered and intensity normalized atlases. Each location in the synthesized image is a weighted average of the registered atlases; atlas weights vary spatially. The weights are determined using the joint label fusion (JLF) framework. The primary methodological contribution is the application of JLF to MRI signal directly rather than labels. Synthesized images are then used as additional features in a lesion segmentation task using the OASIS classifier, a logistic regression model on intensities from multiple modalities. The addition of JIF synthesized images improved the Dice-Sorensen coefficient (relative to manually drawn gold standards) of lesion segmentations over the standard model segmentations by 0.0462 ± 0.0050 (mean ± standard deviation) at optimal threshold over all subjects and 10 separate training/testing folds.

ADAPTIVE GRADIENT DESCENT OPTIMIZATION OF INITIAL MOMENTA FOR GEODESIC SHOOTING IN DIFFEOMORPHISMS.

Diffeomorphic image registration algorithms are widely used in medical imaging, and require optimization of a high-dimensional nonlinear objective function. The function being optimized has many characteristics that are relevant for optimization but are typically not well understood. Due to that complexity, most authors have used a simple gradient descent, but it is not often discussed how step sizes are chosen or if line searches are used. Further, if a system is to be robust to a range of input images, that may differ to varying degrees, the optimization must be adaptable. Here, we present two methods of adaptable gradient descent with line searches, and test how they affect image registration. The optimization schemes are deployed for geodesic shooting in diffeomorphisms - an approach that is used to quantify anatomical changes, such as atrophy, in longitudinal image pairs. We evaluate the optimization schemes on their convergence characteristics and based on how well the resulting atrophy scores correlate with diagnostic group and mini mental state exam (MMSE) scores. We find that the Barzilai-Borwein method with a backtracking line search outperforms other optimization schemes in convergence time and adaptability by a wide margin. We also find that the variable optimization schemes do not significantly affect the ability to measure atrophy with clinical significance.

THE IMPACT OF MATCHING FUNCTIONAL ON ATROPHY MEASUREMENT FROM GEODESIC SHOOTING IN DIFFEOMORPHISMS.

Longitudinal registration has been used to map brain atrophy and tissue loss patterns over time, in both healthy and demented subjects. However, we have not seen a thorough application of the geodesic shooting in diffeomorphisms framework for this task. The registration model is complex and several choices must be made that may significantly impact the quality of results. One of these decisions is which image matching functional should drive the registration. We investigate four matching functionals for atrophy quantification using geodesic shooting in diffeomorphisms. We check if the choice of matching functional has an impact on the correlation of atrophy scores with clinical variables. We also check the impact of matching functional choice on estimates of the N80 sample size for hypothetical clinical trials that test for slowing of brain atrophy. We find that the mutual information function, which has primarily been used for linear and multi-modal registration, achieves comparable correlation with clinical variables to other matching functionals while yielding better sample size estimates.

A Transformation Similarity Constraint for Groupwise Nonlinear Registration in Longitudinal Neuro Imaging Studies.

Patients with Alzheimer's disease and other brain disorders often show a similar spatial distribution of volume change throughout the brain over time,1,2 but this information is not yet used in registration algorithms to refine the quantification of change. Here, we develop a mathematical basis to incorporate that prior information into a longitudinal structural neuroimaging study. We modify the canonical minimization problem for non-linear registration to include a term that couples a collection of registrations together to enforce group similarity. More specifically, throughout the computation we maintain a group-level representation of the transformations and constrain updates to individual transformations to be similar to this representation. The derivations necessary to produce the Euler-Lagrange equations for the coupling term are presented and a gradient descent algorithm based on the formulation was implemented. We demonstrate using 57 longitudinal image pairs from the Alzheimer's Disease Neuroimaging Initiative (ADNI) that longitudinal registration with such a groupwise coupling prior is more robust to noise in estimating change, suggesting such change maps may have several important applications.

Simultaneous Longitudinal Registration with Group-Wise Similarity Prior.

Here we present an algorithm for the simultaneous registration of N longitudinal image pairs such that information acquired by each pair is used to constrain the registration of each other pair. More specifically, in the geodesic shooting setting for Large Deformation Diffeomorphic Metric Mappings (LDDMM) an average of the initial momenta characterizing the N transformations is maintained throughout and updates to individual momenta are constrained to be similar to this average. In this way, the N registrations are coupled and explore the space of diffeomorphisms as a group, the variance of which is constrained to be small. Our approach is motivated by the observation that transformations learned from images in the same diagnostic category share characteristics. The group-wise consistency prior serves to strengthen the contribution of the common signal among the N image pairs to the transformation for a specific pair, relative to features particular to that pair. We tested the algorithm on 57 longitudinal image pairs of Alzheimer's Disease patients from the Alzheimer's Disease Neuroimaging Initiative and evaluated the ability of the algorithm to produce momenta that better represent the long-term biological processes occurring in the underlying anatomy. We found that for many image pairs, momenta learned with the group-wise prior better predict a third time point image unobserved in the registration.

A Riemannian Framework for Intrinsic Comparison of Closed Genus-Zero Shapes.

We present a framework for intrinsic comparison of surface metric structures and curvatures. This work parallels the work of Kurtek et al. on parameterization-invariant comparison of genus zero shapes. Here, instead of comparing the embedding of spherically parameterized surfaces in space, we focus on the first fundamental form. To ensure that the distance on spherical metric tensor fields is invariant to parameterization, we apply the conjugation-invariant metric arising from the L2 norm on symmetric positive definite matrices. As a reparameterization changes the metric tensor by a congruent Jacobian transform, this metric perfectly suits our purpose. The result is an intrinsic comparison of shape metric structure that does not depend on the specifics of a spherical mapping. Further, when restricted to tensors of fixed volume form, the manifold of metric tensor fields and its quotient of the group of unitary diffeomorphisms becomes a proper metric manifold that is geodesically complete. Exploiting this fact, and augmenting the metric with analogous metrics on curvatures, we derive a complete Riemannian framework for shape comparison and reconstruction. A by-product of our framework is a near-isometric and curvature-preserving mapping between surfaces. The correspondence is optimized using the fast spherical fluid algorithm. We validate our framework using several subcortical boundary surface models from the ADNI dataset.