Doublets are a characteristic error source in droplet-based single-cell sequencing data where two cells are encapsulated in the same oil emulsion and are tagged with the same cell barcode. Across type doublets manifest as fictitious phenotypes that can be incorrectly interpreted as novel cell types. DoubletDetection present a novel, fast, unsupervised classifier to detect across-type doublets in single-cell RNA-sequencing data that operates on a count matrix and imposes no experimental constraints. This classifier leverages the creation of in silico synthetic doublets to determine which cells in the input count matrix have gene expression that is best explained by the combination of distinct cell types in the matrix. In this notebook, we will illustrate an example workflow for detecting doublets in single-cell RNA-seq count matrices.

Computational methods that model how the gene expression of a cell is influenced by interacting cells are lacking. We present NicheNet, a method that predicts ligand–target links between interacting cells by combining their expression data with prior knowledge of signaling and gene regulatory networks. We applied NicheNet to the tumor and immune cell microenvironment data and demonstrated that NicheNet can infer active ligands and their gene regulatory effects on interacting cells.

The recent development of experimental methods for measuring chromatin state at single-cell resolution has created a need for computational tools capable of analyzing these datasets. Here we developed Signac, a framework for the analysis of single-cell chromatin data, as an extension of the Seurat R toolkit for single-cell multimodal analysis. **Signac** enables an end-to-end analysis of single-cell chromatin data, including peak calling, quantification, quality control, dimension reduction, clustering, integration with single-cell gene expression datasets, DNA motif analysis, and interactive visualization. Furthermore, Signac facilitates the analysis of multimodal single-cell chromatin data, including datasets that co-assay DNA accessibility with gene expression, protein abundance, and mitochondrial genotype. We demonstrate scaling of the Signac framework to datasets containing over 700,000 cells.