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Xenium Punkst Workflows

This repository provides an open-source, reproducible workflow for analyzing 10x Xenium spatial transcriptomics data using segmentation-free spatial factorization (FICTURE / punkst), with extensions for ROI-based transcript cropping.

The project combines:

  • interactive transcript-level ROI cropping using Napari
  • integration with punkst / FICTURE for segmentation-free modeling
  • orchestration of single-sample and multi-sample analyses
  • documentation designed for reproducibility and academic reuse

This repository is intended to serve as both a research pipeline and a citable software artifact.


Overview

Xenium datasets often contain millions of transcripts spanning large tissue sections. While whole-slide analysis is valuable, many biological questions focus on specific anatomical regions or require joint analysis across multiple samples.

This workflow supports:

  • region-of-interest (ROI)–focused analysis
  • transcript-level filtering without cell segmentation
  • consistent analysis across many Xenium samples
  • scalable workflows for large spatial datasets

A high-level conceptual description is provided in docs/overview.md.


Key components

ROI-based transcript cropping (cropping/)

  • Interactive visualization and ROI definition using Napari
  • Polygon-based transcript filtering
  • Deterministic cropping given the same polygon
  • Outputs compatible with downstream punkst / FICTURE analysis

See cropping/README.md for details.


Segmentation-free spatial factorization (punkst/)

  • punkst / FICTURE is included as a Git submodule
  • Upstream code is not copied or modified directly
  • Authorship, licensing, and versioning are preserved

Original repository: https://github.com/yichen-si/punkst

This repository contributes workflows and orchestration around FICTURE, not a reimplementation of the method itself.


Analysis workflows (workflows/)

  • Single-sample workflows
  • Multi-sample joint analysis workflows
  • SVG flowcharts suitable for presentations and posters

See workflows/README.md.


Reproducibility

Reproducibility considerations are documented in docs/reproducibility.md, including:

  • deterministic vs interactive steps
  • ROI provenance
  • submodule versioning
  • environment considerations

Zenodo is used to archive tagged releases and provide a DOI.


Citation

If you use this workflow, please cite:

Sabir Y, Huang M, Cruz-Martín A.
Xenium ROI Cropping + FICTURE Workflows.
Zenodo. https://doi.org/10.5281/zenodo.18490063

Please also cite the original FICTURE publication when using downstream factorization.


License

This project is released under the MIT License.

See the LICENSE file for details.


Notes

  • Large Xenium datasets are not included in this repository.
  • This workflow is designed for research and exploratory analysis.
  • Contributions and issues are welcome via GitHub.

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