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Introduction

Spatial transcriptomics spans a range of technologies – from spot-based platforms like Visium and Slide-seq to subcellular-resolution methods like Xenium and MERFISH. scConvert handles spatial data from any platform that produces a Seurat object or h5ad file, preserving coordinates, images, and metadata through format conversion.

Supported spatial technologies

Technology Resolution Coordinates Images h5ad roundtrip
10x Visium ~55 um spots Gridded H&E tissue Full support
10x Visium HD 2–8 um bins Dense grid H&E tissue Coordinates + image
Slide-seq v2 ~10 um beads Continuous None Coordinates
10x Xenium Subcellular Molecule-based DAPI/IF Coordinates via FOV
MERFISH (Vizgen) Subcellular Molecule-based DAPI Coordinates via FOV
CosMx (NanoString) Subcellular Molecule-based Morphology Coordinates via FOV
CODEX (Akoya) Cell-level Centroids Fluorescence Coordinates
Stereo-seq (BGI) ~0.5 um Continuous H&E Coordinates

All technologies store spatial coordinates in the h5ad obsm/spatial field. Visium additionally stores tissue images and scale factors in uns/spatial.

Visium example: mouse brain

We demonstrate a full roundtrip with the shipped Visium demo dataset (400 mouse brain spots, 1,500 genes, 15 clusters).

spatial_path <- system.file("extdata", "spatial_demo.rds", package = "scConvert")
brain <- readRDS(spatial_path)

cat("Spots:", ncol(brain), "\n")
#> Spots: 400
cat("Genes:", nrow(brain), "\n")
#> Genes: 1500
cat("Image:", Images(brain), "\n")
#> Image: anterior1
cat("Assay:", Assays(brain), "\n")
#> Assay: Spatial

Spatial gene expression

Ttr (Transthyretin) is expressed in the choroid plexus and shows strong spatial localization.

SpatialFeaturePlot(brain, features = "Ttr", pt.size.factor = 1.6) +
  ggplot2::ggtitle("Ttr expression (original)")

Convert and load back

h5ad_file <- tempfile(fileext = ".h5ad")
writeH5AD(brain, h5ad_file, overwrite = TRUE)
brain_rt <- readH5AD(h5ad_file, verbose = FALSE)

cat("Roundtrip spots:", ncol(brain_rt), "\n")
#> Roundtrip spots: 400
cat("Roundtrip genes:", nrow(brain_rt), "\n")
#> Roundtrip genes: 1500
cat("Image preserved:", length(Images(brain_rt)) > 0, "\n")
#> Image preserved: TRUE

Verify coordinate preservation

cat("Barcodes match:", all(colnames(brain) == colnames(brain_rt)), "\n")
#> Barcodes match: TRUE
cat("Features match:", all(rownames(brain) == rownames(brain_rt)), "\n")
#> Features match: TRUE
cat("Clusters match:",
    all(as.character(brain$seurat_clusters) ==
        as.character(brain_rt$seurat_clusters)), "\n")
#> Clusters match: TRUE

Clusters preserved through roundtrip

DimPlot(brain_rt, reduction = "umap", group.by = "seurat_clusters",
        label = TRUE, repel = TRUE) +
  ggplot2::ggtitle("UMAP clusters (after h5ad roundtrip)")

Another gene: Nrgn

Nrgn (Neurogranin) is a cortical neuron marker with a different spatial pattern than Ttr.

FeaturePlot(brain_rt, features = "Nrgn", reduction = "umap") +
  ggplot2::ggtitle("Nrgn expression (UMAP, after roundtrip)")

Working with other spatial platforms

For any h5ad file with spatial coordinates, the same workflow applies:

# Load spatial h5ad from any platform
obj <- readH5AD("merfish_data.h5ad")

# Check what was detected
cat("Images:", Images(obj), "\n")

# Convert to another format
scConvert("merfish_data.h5ad", "merfish_data.h5seurat")

Technologies without tissue images (Slide-seq, MERFISH, CODEX) store only coordinates in obsm/spatial. scConvert reads these into the Seurat object and makes them available for plotting and downstream analysis.

Python interop

# Requires Python with scanpy installed
import scanpy as sc

adata = sc.read_h5ad("brain.h5ad")
print(adata)

# Works with any spatial technology
sc.pl.spatial(adata, color="Ttr")  # Visium (with image)
sc.pl.embedding(adata, basis="spatial", color="cluster")  # Generic spatial

Cleanup

Session Info

sessionInfo()
#> R version 4.5.2 (2025-10-31)
#> Platform: aarch64-apple-darwin20
#> Running under: macOS Tahoe 26.3
#> 
#> Matrix products: default
#> BLAS:   /System/Library/Frameworks/Accelerate.framework/Versions/A/Frameworks/vecLib.framework/Versions/A/libBLAS.dylib 
#> LAPACK: /Library/Frameworks/R.framework/Versions/4.5-arm64/Resources/lib/libRlapack.dylib;  LAPACK version 3.12.1
#> 
#> locale:
#> [1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8
#> 
#> time zone: America/Indiana/Indianapolis
#> tzcode source: internal
#> 
#> attached base packages:
#> [1] stats     graphics  grDevices utils     datasets  methods   base     
#> 
#> other attached packages:
#> [1] ggplot2_4.0.2      Seurat_5.4.0       SeuratObject_5.3.0 sp_2.2-1          
#> [5] scConvert_0.1.0   
#> 
#> loaded via a namespace (and not attached):
#>   [1] RColorBrewer_1.1-3     jsonlite_2.0.0         magrittr_2.0.4        
#>   [4] spatstat.utils_3.2-2   farver_2.1.2           rmarkdown_2.30        
#>   [7] fs_1.6.7               ragg_1.5.0             vctrs_0.7.1           
#>  [10] ROCR_1.0-12            spatstat.explore_3.7-0 htmltools_0.5.9       
#>  [13] sass_0.4.10            sctransform_0.4.3      parallelly_1.46.1     
#>  [16] KernSmooth_2.23-26     bslib_0.10.0           htmlwidgets_1.6.4     
#>  [19] desc_1.4.3             ica_1.0-3              plyr_1.8.9            
#>  [22] plotly_4.12.0          zoo_1.8-15             cachem_1.1.0          
#>  [25] igraph_2.2.2           mime_0.13              lifecycle_1.0.5       
#>  [28] pkgconfig_2.0.3        Matrix_1.7-4           R6_2.6.1              
#>  [31] fastmap_1.2.0          MatrixGenerics_1.22.0  fitdistrplus_1.2-6    
#>  [34] future_1.69.0          shiny_1.13.0           digest_0.6.39         
#>  [37] S4Vectors_0.48.0       patchwork_1.3.2        tensor_1.5.1          
#>  [40] RSpectra_0.16-2        irlba_2.3.7            GenomicRanges_1.62.1  
#>  [43] textshaping_1.0.4      labeling_0.4.3         progressr_0.18.0      
#>  [46] spatstat.sparse_3.1-0  httr_1.4.8             polyclip_1.10-7       
#>  [49] abind_1.4-8            compiler_4.5.2         bit64_4.6.0-1         
#>  [52] withr_3.0.2            S7_0.2.1               fastDummies_1.7.5     
#>  [55] MASS_7.3-65            tools_4.5.2            lmtest_0.9-40         
#>  [58] otel_0.2.0             httpuv_1.6.16          future.apply_1.20.2   
#>  [61] goftest_1.2-3          glue_1.8.0             nlme_3.1-168          
#>  [64] promises_1.5.0         grid_4.5.2             Rtsne_0.17            
#>  [67] cluster_2.1.8.2        reshape2_1.4.5         generics_0.1.4        
#>  [70] hdf5r_1.3.12           gtable_0.3.6           spatstat.data_3.1-9   
#>  [73] tidyr_1.3.2            data.table_1.18.2.1    XVector_0.50.0        
#>  [76] BiocGenerics_0.56.0    BPCells_0.2.0          spatstat.geom_3.7-0   
#>  [79] RcppAnnoy_0.0.23       ggrepel_0.9.7          RANN_2.6.2            
#>  [82] pillar_1.11.1          stringr_1.6.0          spam_2.11-3           
#>  [85] RcppHNSW_0.6.0         later_1.4.8            splines_4.5.2         
#>  [88] dplyr_1.2.0            lattice_0.22-9         survival_3.8-6        
#>  [91] bit_4.6.0              deldir_2.0-4           tidyselect_1.2.1      
#>  [94] miniUI_0.1.2           pbapply_1.7-4          knitr_1.51            
#>  [97] gridExtra_2.3          Seqinfo_1.0.0          IRanges_2.44.0        
#> [100] scattermore_1.2        stats4_4.5.2           xfun_0.56             
#> [103] matrixStats_1.5.0      UCSC.utils_1.6.1       stringi_1.8.7         
#> [106] lazyeval_0.2.2         yaml_2.3.12            evaluate_1.0.5        
#> [109] codetools_0.2-20       tibble_3.3.1           cli_3.6.5             
#> [112] uwot_0.2.4             xtable_1.8-8           reticulate_1.45.0     
#> [115] systemfonts_1.3.1      jquerylib_0.1.4        GenomeInfoDb_1.46.2   
#> [118] dichromat_2.0-0.1      Rcpp_1.1.1             globals_0.19.1        
#> [121] spatstat.random_3.4-4  png_0.1-8              spatstat.univar_3.1-6 
#> [124] parallel_4.5.2         pkgdown_2.2.0          dotCall64_1.2         
#> [127] listenv_0.10.1         viridisLite_0.4.3      scales_1.4.0          
#> [130] ggridges_0.5.7         purrr_1.2.1            crayon_1.5.3          
#> [133] rlang_1.1.7            cowplot_1.2.0