Single-cell RNA sequencing (scRNA-seq) data often encountered technical artifacts called "doublets" which are two cells that are sequenced under the same cellular barcode.
Doublets formed from different cell types or states are called heterotypic and homotypic otherwise. These factors constrain cell throughput and may result in misleading biological interpretations.
DoubletFinder (McGinnis, Murrow, and Gartner 2019) is one of the methods proposed for doublet detection. In this notebook, we will illustrate an example workflow of DoubletFinder. We use a 10x Genomics dataset which captures peripheral blood mononuclear cells (PBMCs) from a healthy donor stained with a panel of 31 TotalSeq™-B antibodies (BioLegend).
PopV uses popular vote of a variety of cell-type transfer tools to classify cell-types in a query dataset based on a test dataset.
Using this variety of algorithms, they compute the agreement between those algorithms and use this agreement to predict which cell-types have a high likelihood of the same cell-types observed in the reference.
Single-cell RNA sequencing (scRNA-seq) data have allowed us to investigate cellular heterogeneity and the kinetics of a biological process. Some studies need to understand how cells change state, and corresponding genes during the process, but it is challenging to track the cell development in scRNA-seq protocols. Therefore, a variety of statistical and computational methods have been proposed for lineage inference (or pseudotemporal ordering) to reconstruct the states of cells according to the developmental process from the measured snapshot data. Specifically, lineage refers to an ordered transition of cellular states, where individual cells represent points along. pseudotime is a one-dimensional variable representing each cell’s transcriptional progression toward the terminal state.
Slingshot which is one of the methods suggested for lineage reconstruction and pseudotime inference from single-cell gene expression data. In this notebook, we will illustrate an example workflow for cell lineage and pseudotime inference using Slingshot. The notebook is inspired by Slingshot's vignette and modified to demonstrate how the tool works on BioTuring's platform.
CellRank2 (Weiler et al, 2023) is a powerful framework for studying cellular fate using single-cell RNA sequencing data. It can handle millions of cells and different data types efficiently. This tool can identify cell fate and probabilities across various data sets. It also allows for analyzing transitions over time and uncovering key genes in developmental processes. Additionally, CellRank2 estimates cell-specific transcription and degradation rates, aiding in understanding differentiation trajectories and regulatory mechanisms.
In this notebook, we will use a primary tumor sample of patient T71 from the dataset GSE137804 (Dong R. et al, 2020) as an example. We have performed RNA-velocity analysis and pseudotime calculation on this dataset in scVelo (Bergen et al, 2020) notebook. The output will be then loaded into this CellRank2 notebook for further analysis.
This notebook is based on the tutorial provided on CellRank2 documentation. We have modified the notebook and changed the input data to show how the tool works on BioTuring's platform.
BPCells is a package for high performance single cell analysis on RNA-seq and ATAC-seq datasets. It can analyze a 1.3M cell dataset with 2GB of RAM in under 10 minutes. This makes analysis of million-cell datasets practical on a laptop.
BPCells provides:
* Efficient storage of single cell datasets via bitpacking compression
* Fast, disk-backed RNA-seq and ATAC-seq data processing powered by C++
* Downstream analysis such as marker genes, and clustering
* Interoperability with AnnData, 10x datasets, R sparse matrices, and GRanges