BrainScope: interactive visual exploration of the spatial and temporal human brain transcriptome

Spatial and temporal brain transcriptomics has recently emerged as an invaluable data source for molecular neuroscience. The complexity of such data poses considerable challenges for analysis and visualization. We present BrainScope: a web portal for fast, interactive visual exploration of the Allen Atlases of the adult and developing human brain transcriptome. Through a novel methodology to explore high-dimensional data (dual t-SNE), BrainScope enables the linked, all-in-one visualization of genes and samples across the whole brain and genome, and across developmental stages. We show that densities in t-SNE scatter plots of the spatial samples coincide with anatomical regions, and that densities in t-SNE scatter plots of the genes represent gene co-expression modules that are significantly enriched for biological functions. We also show that the topography of the gene t-SNE maps reflect brain region-specific gene functions, enabling hypothesis and data driven research. We demonstrate the discovery potential of BrainScope through three examples: (i) analysis of cell type specific gene sets, (ii) analysis of a set of stable gene co-expression modules across the adult human donors and (iii) analysis of the evolution of co-expression of oligodendrocyte specific genes over developmental stages. BrainScope is publicly accessible at www.brainscope.nl.FSW – Publicaties zonder aanstelling Universiteit Leide

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion

Comparative cellular analysis of motor cortex in human, marmoset and mouse

The primary motor cortex (M1) is essential for voluntary fine-motor control and is functionally conserved across mammals1. Here, using high-throughput transcriptomic and epigenomic profiling of more than 450,000 single nuclei in humans, marmoset monkeys and mice, we demonstrate a broadly conserved cellular makeup of this region, with similarities that mirror evolutionary distance and are consistent between the transcriptome and epigenome. The core conserved molecular identities of neuronal and non-neuronal cell types allow us to generate a cross-species consensus classification of cell types, and to infer conserved properties of cell types across species. Despite the overall conservation, however, many species-dependent specializations are apparent, including differences in cell-type proportions, gene expression, DNA methylation and chromatin state. Few cell-type marker genes are conserved across species, revealing a short list of candidate genes and regulatory mechanisms that are responsible for conserved features of homologous cell types, such as the GABAergic chandelier cells. This consensus transcriptomic classification allows us to use patch-seq (a combination of whole-cell patch-clamp recordings, RNA sequencing and morphological characterization) to identify corticospinal Betz cells from layer 5 in non-human primates and humans, and to characterize their highly specialized physiology and anatomy. These findings highlight the robust molecular underpinnings of cell-type diversity in M1 across mammals, and point to the genes and regulatory pathways responsible for the functional identity of cell types and their species-specific adaptations

Overview + detail visualization for ensembles of diffusion tensors

A Diffusion Tensor Imaging (DTI) group study consists of a collection of volumetric diffusion tensor datasets (i.e., an ensemble) acquired from a group of subjects. The multivariate nature of the diffusion tensor imposes challenges on the analysis and the visualization. These challenges are commonly tackled by reducing the diffusion tensors to scalar-valued quantities that can be analyzed with common statistical tools. However, reducing tensors to scalars poses the risk of losing intrinsic information about the tensor. Visualization of tensor ensemble data without loss of information is still a largely unsolved problem. In this work, we propose an overview + detail visualization to facilitate the tensor ensemble exploration. We define an ensemble representative tensor and variations in terms of the three intrinsic tensor properties (i.e., scale, shape, and orientation) separately. The ensemble summary information is visually encoded into the newly designed aggregate tensor glyph which, in a spatial layout, functions as the overview. The aggregate tensor glyph guides the analyst to interesting areas that would need further detailed inspection. The detail views reveal the original information that is lost during aggregation. It helps the analyst to further understand the sources of variation and formulate hypotheses. To illustrate the applicability of our prototype, we compare with most relevant previous work through a user study and we present a case study on the analysis of a brain diffusion tensor dataset ensemble from healthy volunteers

A Progressive Approach for Uncertainty Visualization in Diffusion Tensor Imaging

Diffusion Tensor Imaging (DTI) is a non-invasive magnetic resonance imaging technique that, combined with fiber tracking algorithms, allows the characterization and visualization of white matter structures in the brain. The resulting fiber tracts are used, for example, in tumor surgery to evaluate the potential brain functional damage due to tumor resection. The DTI processing pipeline from image acquisition to the final visualization is rather complex generating undesirable uncertainties in the final results. Most DTI visualization techniques do not provide any information regarding the presence of uncertainty. When planning surgery, a fixed safety margin around the fiber tracts is often used; however, it cannot capture local variability and distribution of the uncertainty, thereby limiting the informed decision-making process. Stochastic techniques are a possibility to estimate uncertainty for the DTI pipeline. However, it has high computational and memory requirements that make it infeasible in a clinical setting. The delay in the visualization of the results adds hindrance to the workflow. We propose a progressive approach that relies on a combination of wild-bootstrapping and fiber tracking to be used within the progressive visual analytics paradigm. We present a local bootstrapping strategy, which reduces the computational and memory costs, and provides fiber-tracking results in a progressive manner. We have also implemented a progressive aggregation technique that computes the distances in the fiber ensemble during progressive bootstrap computations. We present experiments with different scenarios to highlight the benefits of using our progressive visual analytic pipeline in a clinical workflow along with a use case and analysis obtained by discussions with our collaborators.Comp Graphics & Visualisatio

Uncertainty in the DTI Visualization Pipeline

Diffusion-Weighted Magnetic Resonance Imaging (DWI) enables the in-vivo visualization of fibrous tissues such as white matter in the brain. Diffusion-Tensor Imaging (DTI) specifically models the DWI diffusion measurements as a second order-tensor. The processing pipeline to visualize this data, from image acquisition to the final rendering, is rather complex. It involves a considerable amount of measurements, parameters and model assumptions, all of which generate uncertainties in the final result which typically are not shown to the analyst in the visualization. In recent years, there has been a considerable amount of work on the visualization of uncertainty in DWI, and specifically DTI. In this chapter, we primarily focus on DTI given its simplicity and applicability, however, several aspects presented are valid for DWI as a whole. We explore the various sources of uncertainties involved, approaches for modeling those uncertainties, and, finally, we survey different strategies to visually represent them. We also look at several related methods of uncertainty visualization that have been applied outside DTI and discuss how these techniques can be adopted to the DTI domain. We conclude our discussion with an overview of potential research directions.Comp Graphics & Visualisatio

Focus+context exploration of hierarchical embeddings

Hierarchical embeddings, such as HSNE, address critical visual and computational scalability issues of traditional techniques for dimensionality reduction. The improved scalability comes at the cost of the need for increased user interaction for exploration. In this paper, we provide a solution for the interactive visual Focus+Context exploration of such embeddings. We explain how to integrate embedding parts from different levels of detail, corresponding to focus and context groups, in a joint visualization. We devise an according interaction model that relates typical semantic operations on a Focus+Context visualization with the according changes in the level-of-detail-hierarchy of the embedding, including also a mode for comparative Focus+Context exploration and extend HSNE to incorporate the presented interaction model. In order to demonstrate the effectiveness of our approach, we present a use case based on the visual exploration of multi-dimensional images.</p

ImaCytE: Visual Exploration of Cellular Micro-Environments for Imaging Mass Cytometry Data

Tissue functionality is determined by the characteristics of tissue-resident cells and their interactions within their microenvironment. Imaging Mass Cytometry offers the opportunity to distinguish cell types with high precision and link them to their spatial location in intact tissues at sub-cellular resolution. This technology produces large amounts of spatially-resolved high-dimensional data, which constitutes a serious challenge for the data analysis. We present an interactive visual analysis workflow for the end-to-end analysis of Imaging Mass Cytometry data that was developed in close collaboration with domain expert partners. We implemented the presented workflow in an interactive visual analysis tool; ImaCytE. Our workflow is designed to allow the user to discriminate cell types according to their protein expression profiles and analyze their cellular microenvironments, aiding in the formulation or verification of hypotheses on tissue architecture and function. Finally, we show the effectiveness of our workflow and ImaCytE through a case study performed by a collaborating specialist

Incorporating Texture Information into Dimensionality Reduction for High-Dimensional Images

High-dimensional imaging is becoming increasingly relevant in many fields from astronomy and cultural heritage to systems biology. Visual exploration of such high-dimensional data is commonly facilitated by dimensionality reduction. However, common dimensionality reduction methods do not include spatial information present in images, such as local texture features, into the construction of low-dimensional embeddings. Consequently, exploration of such data is typically split into a step focusing on the attribute space followed by a step focusing on spatial information, or vice versa. In this paper, we present a method for incorporating spatial neighborhood information into distance-based dimensionality reduction methods, such as t-Distributed Stochastic Neighbor Embedding (t-SNE). We achieve this by modifying the distance measure between high-dimensional attribute vectors associated with each pixel such that it takes the pixel's spatial neighborhood into account. Based on a classification of different methods for comparing image patches, we explore a number of different approaches. We compare these approaches from a theoretical and experimental point of view. Finally, we illustrate the value of the proposed methods by qualitative and quantitative evaluation on synthetic data and two real-world use cases.</p