Background
Methods
Patient collective
Immunohistochemistry staining
Comment on image ethics
Image acquisition and pre-processing (Additional file 1: Figure S1)
Definition and segmentation of spaces and niches
Image processing in MATLAB to detect cells (Additional file 1: Figure S1)
Introduction of a field (Additional file 1: Figure S1)
Normalization of field values
Calculation of the gradient and the divergence
Statistical evaluation (Additional file 1: Figure S1)
Results
Proof of principle for the description of colocalization and possible spatial interaction via a scalar field
Comparison to colocalization via colour channel analysis
Proof of principle for valid distance measurements through histogram intersection
Proof of principle for description and detection of object clusters via calculation of the gradient and the divergence
Application on histological images
Example 1: Spatial interaction of B-cell markers in a focal, dense infiltration
Example 2: Allocation and spatial interaction of B- and T-cells in a focal, dense infiltration
Example 3: Co-localization of T-cell markers in a lose infiltration
Direct interaction/RBFdirect
| Indirect interaction/RBFindirect
| |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
CD3 | CD3 Bone | CD4 | CD4 Bone | CD8 | CD8 Bone | CD20 | CD20 Bone | CD3 | CD3 Bone | CD4 | CD4 Bone | CD8 | CD8 Bone | CD20 | CD20 Bone | |
CD3 | MCD3 = 0.00 | MCD3 = 0.00b
| MCD3 = 0.02 | MCD3 = 0.00b
| MCD3 = 0.02 | MCD3 = 0.10a
| MCD3 = 0.00 | MCD3 = 0.60 | MCD3 = 0.54 | MCD3 = 0.36 | MCD3 = 0.54 | MCD3 = 0.36 | MCD3 = 0.69a
| MCD3 = 0.60 | ||
CD3 Bone | MCD3Bone = 0.00 | MCD3Bone = 0.03 | MCD3Bone = 0.43a
| MCD3Bone = 0.00 | MCD3Bone = 0.43b
| MCD3Bone=0.00 | MCD3Bone=0.88 | MCD3Bone = 0.89 | MCD3Bone = 0.71 | MCD3Bone = 0.50 | MCD3Bone = 0.71 | MCD3Bone = 0.50 | MCD3Bone=0.59 | MCD3Bone=0.98 | ||
CD4 | MCD4 = 0.00b
| MCD4 = 0.61 | MCD4 = 0.00 | MCD4 = 0.00 | MCD4 = 0.00 | MCD4 = 0.00 | MCD4 = 0.39 | MCD4 = 0.91 | MCD4 = 0.80 | MCD4 = 0.24 | MCD4 = 0.72 | MCD4 = 0.21 | MCD4 = 0.69 | MCD4 = 0.24 | ||
CD4 Bone | MCD4Bone = 0.00 | MCD4Bone = 0.79a
| MCD4Bone = 0.00 | MCD4Bone = 0.00 | MCD4Bone=0.84 | MCD4Bone=0.01 | MCD4Bone=0.31 | MCD4Bone = 0.88 | MCD4Bone = 0.83 | MCD4Bone = 0.36 | MCD4Bone = 0.66 | MCD4Bone=0.92 | MCD4Bone=0.54 | MCD4Bone=0.82 | ||
CD8 | MCD8 = 0.00b
| MCD8 = 0.00 | MCD8 =0.00 | MCD8 = 0.00 | MCD8 = 0.00 | MCD8 = 0.00 | MCD8 = 0.00 | MCD8 = 0.91 | MCD8 = 0.80 | MCD8 = 0.50 | MCD8 = 0.31 | MCD8 = 0.27 | MCD8 = 0.80 | MCD8 = 0.51 | ||
CD8 Bone | MCD8Bone = 0.00 | MCD8Bone = 0.71b
| MCD8Bone = 0.00 | MCD8Bone = 0.96 | MCD8Bone = 0.00 | MCD8Bone=0.01 | MCD8Bone=0.25 | MCD8Bone = 0.89 | MCD8Bone = 0.83 | MCD8Bone = 0.21 | MCD8Bone = 1.00 | MCD8Bone = 0.64 | MCD8Bone=0.51 | MCD8Bone=0.83 | ||
CD20 | MCD20=0.15
a
| MCD20=0.00
| MCD20 = 0.00 | MCD20 = 0.06 | MCD20 = 0.00 | MCD20 = 0.07 | MCD20 = 0.00 | MCD20=0.99
a
| MCD20=0.57
| MCD20 = 0.59 | MCD20 = 0.31 | MCD20 = 0.99 | MCD20 = 0.27 | MCD20 = 0.58 | ||
CD20 Bone | MCD20Bone=0.00
| MCD20Bone=0.91
| MCD20Bone=0.03 | MCD20Bone=0.46 | MCD20Bone=0.00 | MCD20Bone=0.44 | MCD20Bone=0.00 | MCD20Bone=0.90
| MCD20Bone=0.99
| MCD20Bone=0.36 | MCD20Bone=0.50 | MCD20Bone=0.68 | MCD20Bone=0.46 | MCD20Bone=0.60 |