bio-workflows-spatial-pipeline

Version Compatibility

Reference examples tested with: matplotlib 3.8+, numpy 1.26+, scanpy 1.10+, squidpy 1.3+

Before using code patterns, verify installed versions match. If versions differ:

• Python: pip show <package> then help(module.function) to check signatures

If code throws ImportError, AttributeError, or TypeError, introspect the installed package and adapt the example to match the actual API rather than retrying.

Spatial Transcriptomics Pipeline

"Analyze my spatial transcriptomics data end-to-end" → Orchestrate data loading (squidpy/scanpy), QC, normalization, spatial domain detection, deconvolution (cell2location), spatial neighbor analysis, cell-cell communication, and tissue visualization.

Complete workflow for analyzing Visium, Xenium, or other spatial transcriptomics data.

Workflow Overview

Spatial data (Space Ranger output)
    |
    v
[1. Load Data] ---------> Read Visium/Xenium
    |
    v
[2. QC & Preprocessing] -> Filter, normalize
    |
    v
[3. Clustering] --------> Standard scRNA-seq clustering
    |
    v
[4. Spatial Analysis] --> Neighbors, statistics
    |
    v
[5. Domain Detection] --> Spatial domains
    |
    v
[6. Visualization] -----> Spatial plots
    |
    v
Annotated spatial data

Primary Path: Squidpy + Scanpy

Step 1: Load Data

import scanpy as sc
import squidpy as sq
import numpy as np
import matplotlib.pyplot as plt

# Load Visium data (Space Ranger output)
adata = sq.read.visium('spaceranger_output/')

# Or load from specific files
adata = sc.read_10x_h5('filtered_feature_bc_matrix.h5')
adata.uns['spatial'] = ...  # Add spatial info

# For Xenium
adata = sq.read.xenium('xenium_output/')

print(f'Loaded: {adata.n_obs} spots/cells, {adata.n_vars} genes')

Step 2: Quality Control

# QC metrics
adata.var['mt'] = adata.var_names.str.startswith('MT-')
sc.pp.calculate_qc_metrics(adata, qc_vars=['mt'], inplace=True)

# Visualize QC
fig, axes = plt.subplots(1, 3, figsize=(15, 4))
sc.pl.spatial(adata, color='total_counts', ax=axes[0], show=False)
sc.pl.spatial(adata, color='n_genes_by_counts', ax=axes[1], show=False)
sc.pl.spatial(adata, color='pct_counts_mt', ax=axes[2], show=False)
plt.savefig('qc_spatial.pdf')

# Filter
sc.pp.filter_cells(adata, min_counts=500)
sc.pp.filter_cells(adata, min_genes=200)
sc.pp.filter_genes(adata, min_cells=10)
adata = adata[adata.obs.pct_counts_mt < 25, :]

print(f'After QC: {adata.n_obs} spots/cells')