Benchmarking cell type annotation methods for 10x Xenium spatial transcriptomics data

Abstract Background Imaging-based spatial transcriptomics technologies allow us to explore spatial gene expression profiles at the cellular level. Cell type annotation of imaging-based spatial data is challenging due to the small gene panel, but it is a crucial step for downstream analyses. Many goo...

Full description

Saved in:
Bibliographic Details
Main Authors: Jinming Cheng, Xinyi Jin, Gordon K. Smyth, Yunshun Chen
Format: Article
Language:English
Published: BMC 2025-01-01
Series:BMC Bioinformatics
Subjects:
Online Access:https://doi.org/10.1186/s12859-025-06044-0
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Abstract Background Imaging-based spatial transcriptomics technologies allow us to explore spatial gene expression profiles at the cellular level. Cell type annotation of imaging-based spatial data is challenging due to the small gene panel, but it is a crucial step for downstream analyses. Many good reference-based cell type annotation tools have been developed for single-cell RNA sequencing and sequencing-based spatial transcriptomics data. However, the performance of the reference-based cell type annotation tools on imaging-based spatial transcriptomics data has not been well studied yet. Results We compared performance of five reference-based methods (SingleR, Azimuth, RCTD, scPred and scmapCell) with the marker-gene-based manual annotation method on an imaging-based Xenium data of human breast cancer. A practical workflow has been demonstrated for preparing a high-quality single-cell RNA reference, evaluating the accuracy, and estimating the running time for reference-based cell type annotation tools. Conclusions SingleR was the best performing reference-based cell type annotation tool for the Xenium platform, being fast, accurate and easy to use, with results closely matching those of manual annotation.
ISSN:1471-2105