Image Analysis for Spatial Transcriptomics
Extract features and segment tissue images in spatial transcriptomics data.
Required Imports
python
import squidpy as sq import scanpy as sc import numpy as np import matplotlib.pyplot as plt from skimage import io, filters, segmentation
Access Tissue Images
python
# Get image from Visium data
library_id = list(adata.uns['spatial'].keys())[0]
img_dict = adata.uns['spatial'][library_id]['images']
# High and low resolution images
hires = img_dict['hires']
lowres = img_dict['lowres']
print(f'Hires shape: {hires.shape}')
print(f'Lowres shape: {lowres.shape}')
# Get scale factors
scalef = adata.uns['spatial'][library_id]['scalefactors']
spot_diameter = scalef['spot_diameter_fullres']
hires_scale = scalef['tissue_hires_scalef']
Create ImageContainer
python
# Squidpy's ImageContainer for organized image handling
img = sq.im.ImageContainer(adata.uns['spatial'][library_id]['images']['hires'])
print(img)
# Or load from file
img = sq.im.ImageContainer('tissue_image.tif')
# Access the image array
arr = img['image'].values
Extract Image Features per Spot
python
# Calculate image features for each spot
sq.im.calculate_image_features(
adata,
img,
features=['summary', 'histogram', 'texture'],
key_added='img_features',
spot_scale=1.0, # Fraction of spot diameter
n_jobs=4,
)
# Features stored in adata.obsm['img_features']
print(f"Image features shape: {adata.obsm['img_features'].shape}")
Available Image Features
python
# Summary statistics
sq.im.calculate_image_features(adata, img, features='summary')
# Mean, std, etc. per channel
# Histogram features
sq.im.calculate_image_features(adata, img, features='histogram', features_kwargs={'histogram': {'bins': 16}})
# Intensity distribution
# Texture features (GLCM)
sq.im.calculate_image_features(adata, img, features='texture')
# Contrast, homogeneity, correlation, ASM
# Custom features
sq.im.calculate_image_features(
adata, img,
features=['summary', 'texture'],
features_kwargs={
'summary': {'quantiles': [0.1, 0.5, 0.9]},
'texture': {'distances': [1, 2], 'angles': [0, np.pi/4, np.pi/2]},
}
)
Segment Cells/Nuclei
python
# Segment using watershed
sq.im.segment(
img,
layer='image',
method='watershed',
channel=0, # Use first channel
thresh=0.5,
)
# Access segmentation mask
seg_mask = img['segmented_watershed'].values
Segment with Cellpose
python
# Cellpose provides better cell segmentation from cellpose import models # Load model model = models.Cellpose(model_type='nuclei') # Get image array image = img['image'].values[:, :, 0] # Single channel # Segment masks, flows, styles, diams = model.eval(image, diameter=30, channels=[0, 0]) # Add to ImageContainer img.add_img(masks, layer='cellpose_masks')
Extract Spot Image Crops
python
# Get image crop around each spot
def get_spot_crop(adata, img_arr, spot_idx, crop_size=100):
coords = adata.obsm['spatial'][spot_idx]
scalef = adata.uns['spatial'][library_id]['scalefactors']['tissue_hires_scalef']
x, y = int(coords[0] * scalef), int(coords[1] * scalef)
half = crop_size // 2
crop = img_arr[max(0, y-half):y+half, max(0, x-half):x+half]
return crop
# Get crop for spot 0
crop = get_spot_crop(adata, hires, 0)
plt.imshow(crop)
Color Deconvolution (H&E)
python
from skimage.color import rgb2hed, hed2rgb
# Separate H&E stains
hed = rgb2hed(hires)
hematoxylin = hed[:, :, 0]
eosin = hed[:, :, 1]
dab = hed[:, :, 2]
# Visualize
fig, axes = plt.subplots(1, 3, figsize=(15, 5))
axes[0].imshow(hematoxylin, cmap='gray')
axes[0].set_title('Hematoxylin')
axes[1].imshow(eosin, cmap='gray')
axes[1].set_title('Eosin')
axes[2].imshow(hires)
axes[2].set_title('Original')
plt.tight_layout()
Compute Morphological Features
python
from skimage.measure import regionprops_table
# Get properties from segmentation
props = regionprops_table(
seg_mask,
intensity_image=hires[:, :, 0],
properties=['label', 'area', 'eccentricity', 'solidity', 'mean_intensity']
)
import pandas as pd
morph_df = pd.DataFrame(props)
print(morph_df.describe())
Use Image Features for Clustering
python
# Combine expression and image features
import numpy as np
# Get expression PCA
expr_pca = adata.obsm['X_pca'][:, :20]
# Get image features
img_features = adata.obsm['img_features']
# Scale and combine
from sklearn.preprocessing import StandardScaler
expr_scaled = StandardScaler().fit_transform(expr_pca)
img_scaled = StandardScaler().fit_transform(img_features)
# Weight combination
alpha = 0.3 # Image weight
combined = np.hstack([
(1 - alpha) * expr_scaled,
alpha * img_scaled
])
adata.obsm['X_combined'] = combined
# Cluster on combined features
sc.pp.neighbors(adata, use_rep='X_combined')
sc.tl.leiden(adata, key_added='combined_leiden')
Smooth Expression with Image
python
# Use image similarity to smooth expression from scipy.spatial.distance import cdist # Compute image similarity matrix img_features = adata.obsm['img_features'] img_sim = 1 / (1 + cdist(img_features, img_features, metric='euclidean')) # Normalize img_sim = img_sim / img_sim.sum(axis=1, keepdims=True) # Smooth expression X_smoothed = img_sim @ adata.X adata.layers['img_smoothed'] = X_smoothed
Related Skills
- •spatial-data-io - Load spatial data with images
- •spatial-visualization - Visualize images with expression
- •spatial-domains - Use image features for domain detection