We present Cell2Cell, a novel visual analytics approach for quantifying and visualizing networks of cell-cell interactions in three-dimensional (3D) multi-channel cancerous tissue data. By analyzing cellular interactions, biomedical experts can gain a more accurate understanding of the intricate relationships between cancer and immune cells. Recent methods have focused on inferring interaction based on the proximity of cells in low-resolution 2D multi-channel imaging data. By contrast, we analyze cell interactions by quantifying the presence and levels of specific proteins within a tissue sample (protein expressions) extracted from high-resolution 3D multi-channel volume data. Such analyses have a strong exploratory nature and require a tight integration of domain experts in the analysis loop to leverage their deep knowledge. We propose two complementary semi-automated approaches to cope with the increasing size and complexity of the data interactively: On the one hand, we interpret cell-to-cell interactions as edges in a cell graph and analyze the image signal (protein expressions) along those edges, using spatial as well as abstract visualizations. Complementary, we propose a cell-centered approach, enabling scientists to visually analyze polarized distributions of proteins in three dimensions, which also captures neighboring cells with biochemical and cell biological consequences. We evaluate our application in three case studies, where biologists and medical experts use Cell2Cell to investigate tumor micro-environments to identify and quantify T-cell activation in human tissue data. We confirmed that our tool can fully solve the use cases and enables a streamlined and detailed analysis of cell-cell interactions.

Graph spectral clustering is a fundamental technique in data analysis, which utilizes eigenpairs of the Laplacian matrix to partition graph vertices into clusters. However, classical spectral clustering algorithms require eigendecomposition of the Laplacian matrix, which has cubic time complexity. In this work, we describe pass-efficient spectral clustering algorithms that leverage recent advances in randomized eigendecomposition and the structure of the graph vertex-edge matrix. Furthermore, we derive formulas for their efficient implementation. The resulting algorithms have a linear time complexity with respect to the number of vertices and edges and pass over the graph constant times, making them suitable for processing large graphs stored on slow memory. Experiments validate the accuracy and efficiency of the algorithms.

Cancer is a complex cellular ecosystem where malignant cells coexist and interact with immune, stromal and other cells within the tumor microenvironment (TME). Recent technological advancements in spatially resolved multiplexed imaging at single-cell resolution have led to the generation of large-scale and high-dimensional datasets from biological specimens. This underscores the necessity for automated methodologies that can effectively characterize molecular, cellular and spatial properties of TMEs for various malignancies. This study introduces SpatialCells, an open-source software package designed for region-based exploratory analysis and comprehensive characterization of TMEs using multiplexed single-cell data. The source code and tutorials are available at https://semenovlab.github.io/SpatialCells. SpatialCells efficiently streamlines the automated extraction of features from multiplexed single-cell data and can process samples containing millions of cells. Thus, SpatialCells facilitates subsequent association analyses and machine learning predictions, making it an essential tool in advancing our understanding of tumor growth, invasion and metastasis.