Spatial Proteomics Analysis
Data Loading
python
import scimap as sm
import anndata as ad
# Load CODEX/IMC data (cell x marker matrix with spatial coordinates)
adata = ad.read_h5ad('spatial_proteomics.h5ad')
# Required: spatial coordinates in adata.obsm['spatial']
# Required: protein intensities in adata.X
Preprocessing
python
# Log transform intensities sm.pp.log1p(adata) # Rescale markers (0-1 per marker) sm.pp.rescale(adata) # Combat batch correction if multiple FOVs sm.pp.combat(adata, batch_key='fov')
Phenotyping Cells
python
# Manual gating approach
phenotype_markers = {
'T_cell': ['CD3', 'CD45'],
'B_cell': ['CD20', 'CD45'],
'Macrophage': ['CD68', 'CD163'],
'Tumor': ['panCK', 'Ki67']
}
sm.tl.phenotype_cells(adata, phenotype=phenotype_markers,
gate=0.5, label='phenotype')
# Clustering-based phenotyping
sm.tl.cluster(adata, method='leiden', resolution=1.0)
Spatial Analysis
python
# Build spatial neighbors graph
sm.tl.spatial_distance(adata, x_coordinate='X', y_coordinate='Y')
# Neighborhood enrichment
sm.tl.spatial_interaction(adata, phenotype='phenotype',
method='knn', knn=10)
# Spatial clustering (communities of cells)
sm.tl.spatial_cluster(adata, phenotype='phenotype')
Visualization
python
# Spatial scatter plot
sm.pl.spatial_scatterPlot(adata, colorBy='phenotype',
x='X', y='Y', s=5)
# Heatmap of spatial interactions
sm.pl.spatial_interaction(adata)
# Marker expression overlay
sm.pl.image_viewer(adata, markers=['CD3', 'CD20', 'panCK'])
Integration with Transcriptomics
python
import squidpy as sq # If matched spatial transcriptomics available # Transfer labels or integrate modalities sq.gr.spatial_neighbors(adata_protein) sq.gr.spatial_neighbors(adata_rna) # Compare spatial patterns across modalities
Platform-Specific Notes
| Platform | Markers | Resolution | Notes |
|---|---|---|---|
| CODEX | 40-60 | Subcellular | Cyclic staining |
| IMC | 40+ | 1 um | Metal-tagged antibodies |
| MIBI | 40+ | 260 nm | Mass spectrometry |
Related Skills
- •spatial-transcriptomics/spatial-neighbors - Spatial graph construction
- •spatial-transcriptomics/spatial-domains - Domain identification
- •imaging-mass-cytometry/phenotyping - IMC-specific analysis