Classic Visium Branch — Spot-Level 5-Stage Pipeline
Goal
Run the classic Visium spot-level analysis. Spots are 55 μm and cover multiple cells — deconvolution IS required to estimate cell-type composition (unlike Visium HD / Xenium / Atera which are cell-resolved).
Stages
S1 — Load + QC
- Tool chain:
init_spatial_project→load_visium_data→filter_visium_spots - Plots: QC violin plots / spot count distributions before and after filtering
- Results: filtered h5ad, QC summary
- Review focus: are the filtering thresholds (
min_counts,min_genes,pct_mt) reasonable? Is the retained spot count consistent with the expected tissue size?
S2 — Normalize + Cluster
- Tool chain:
normalize_visium→cluster_spatial_data - Plots: UMAP, spatial scatter colored by cluster
- Results: normalized/clustered h5ad
- Review focus: is the number of clusters biologically plausible? Do clusters segregate spatially?
S3 — Spatial Domains + SVG
- Tool chain:
identify_spatial_domains→find_spatially_variable_genes - Plots: spatial domain map, top SVG spatial plots
- Results: domain assignments, SVG ranked CSV
- Review focus: do spatial domains match known tissue architecture? Do top SVGs make biological sense?
S4 — Deconvolution (required for classic Visium)
- Tool chain:
deconvolve_spatial_destvi(preferred) ordeconvolve_spatial_spotlight - Requires reference scRNA-seq h5ad with cell-type labels in
obs - Plots: cell-type proportion spatial maps, proportion stacked bar
- Results: proportions CSV per spot, deconvolved h5ad
- Review focus: are the dominant cell types consistent with the tissue type? Any unexpected cell type dominating?
S5 — Prepare for Shared Downstream
- Ensure spot-level h5ad has cell-type labels (from deconvolution) attached for neighborhood enrichment / CellChat.
- If the user also wants single-cell reconstruction from classic Visium (optional, unusual), mention STIE as the morphological-deconvolution route — but this is NOT the default.
Outputs
results/01_loading/...results/07_deconvolution/(see parent skill conventions)- Final: spot-level h5ad with deconvolution proportions attached
Biological Interpretation
Use the parent skill's interpretation template for each stage. For S4 specifically, cross-check dominant cell types against tissue type (e.g., gastric cancer should show epithelial/tumor cells in tumor regions).
Stop for Review
Present interpretation at every stage using the template from the parent spatial-transcriptomics skill. Wait for 通过 / 调整 / 跳过 before proceeding.


