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Optimizing Xenium In Situ data utility by quality assessment and best-practice analysis workflows

Sergio Marco Salas, Louis B. Kuemmerle, Christoffer Mattsson-Langseth, Sebastian Tismeyer, Christophe Avenel, Taobo Hu, Habib Rehman, Marco Grillo, Paulo Czarnewski, Saga Helgadóttir, Katarína Tiklová, Axel Andersson, Nima Rafati, Maria Chatzinikolaou, Fabian J. Theis, Malte D. Luecken, Carolina Wählby, Naveed Ishaque, Mats Nilsson

2025Nature Methods94 citationsDOIOpen Access PDF

Abstract

The Xenium In Situ platform is a new spatial transcriptomics product commercialized by 10x Genomics, capable of mapping hundreds of genes in situ at subcellular resolution. Given the multitude of commercially available spatial transcriptomics technologies, recommendations in choice of platform and analysis guidelines are increasingly important. Herein, we explore 25 Xenium datasets generated from multiple tissues and species, comparing scalability, resolution, data quality, capacities and limitations with eight other spatially resolved transcriptomics technologies and commercial platforms. In addition, we benchmark the performance of multiple open-source computational tools, when applied to Xenium datasets, in tasks including preprocessing, cell segmentation, selection of spatially variable features and domain identification. This study serves as an independent analysis of the performance of Xenium, and provides best practices and recommendations for analysis of such datasets.

Topics & Concepts

Computer scienceWorkflowPreprocessorBenchmark (surveying)ScalabilityIdentification (biology)Data miningDomain (mathematical analysis)Data scienceComputational biologyArtificial intelligenceBiologyDatabaseCartographyMathematicsBotanyGeographyMathematical analysisSingle-cell and spatial transcriptomicsGene expression and cancer classificationMolecular Biology Techniques and Applications
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