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Automated Quantification of Hematopoietic Cell – Stromal Cell Interactions in Histological Images of Undecalcified Bone
Published on: April 8, 2015
On the representation of cells in bone marrow pathology by a scalar field: propagation through serial sections,
Cleo-Aron Weis1, Benedict Walter Grießmann2, Christoph Scharff2
1Institute of Pathology, University Medical Centre Mannheim, Heidelberg University, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany. Cleo-Aron.Weis@medma.uni-heidelberg.de.
Insights
This study introduces a novel scalar field method for analyzing cellular interactions in bone marrow histology. This approach enables objective, quantifiable colocalization and spatial interaction analysis from serial sections, overcoming limitations of traditional methods.
Area of Science:
- Histology and Cell Biology
- Bioinformatics and Computational Biology
- Biomedical Imaging Analysis
Background:
- Bone marrow analysis is challenging due to cellular heterogeneity and complex immunophenotypes.
- Characterizing cellular interactions in situ requires advanced techniques beyond standard immunohistochemistry.
- Existing methods struggle with precise delineation of functional compartments and cell populations like regulatory T-cells.
Purpose of the Study:
- To develop a novel method for analyzing cellular interactions and colocalization in bone marrow histology using scalar fields.
- To overcome the limitations of traditional immunohistochemical analysis in complex biological tissues.
- To enable objective, quantifiable assessment of spatial relationships between cells and tissue structures.
Main Methods:
- Objects (cells, bone trabeculae) are transformed into scalar fields via convolution of centroids with radial basis functions.
- Registered serial histological sections are used to propagate these scalar fields.
- Summation fields are generated to describe distributed objects and enable analysis of spatial interactions.
Main Results:
- Image registration enables robust colocalization analysis resistant to matching errors and morphological variations.
- Scalar fields quantify spatial interactions, including direct and paracrine cellular contact, based on field shape.
- Field overlap, calculated via histogram intersection, accurately represents spatial distances between objects.
Conclusions:
- The scalar field approach provides an automatic, objective, and quantifiable method for marker colocalization in serial sections, with lower technical requirements than multicolour staining.
- This method facilitates the analysis of diverse spatial interactions (direct/indirect) by considering field characteristics.
- Summation fields offer cluster definitions based on object proximity, enhancing spatial interaction analysis.
Background:
Immunohistochemical analysis of cellular interactions in the bone marrow in situ is demanding, due to its heterogeneous cellular composition, the poor delineation and overlap of functional compartments and highly complex immunophenotypes of several cell populations (e.g. regulatory T-cells) that require immunohistochemical marker sets for unambiguous characterization. To overcome these difficulties, we herein present an approach to describe objects (e.g. cells, bone trabeculae) by a scalar field that can be propagated through registered images of serial histological sections.
Methods:
The transformation of objects within images (e.g. cells) to a scalar field was performed by convolution of the object's centroids with differently formed radial basis function (e.g. for direct or indirect spatial interaction). On the basis of such a scalar field, a summation field described distributed objects within an image.
Results:
After image registration i) colocalization analysis could be performed on basis scalar field, which is propagated through registered images, and - due to the shape of the field - were barely prone to matching errors and morphological changes by different cutting levels; ii) furthermore, depending on the field shape the colocalization measurements could also quantify spatial interaction (e.g. direct or paracrine cellular contact); ii) the field-overlap, which represents the spatial distance, of different objects (e.g. two cells) could be calculated by the histogram intersection.
Conclusions:
The description of objects (e.g. cells, cell clusters, bone trabeculae etc.) as a field offers several possibilities: First, co-localization of different markers (e.g. by immunohistochemical staining) in serial sections can be performed in an automatic, objective and quantifiable way. In contrast to multicolour staining (e.g. 10-colour immunofluorescence) the financial and technical requirements are fairly minor. Second, the approach allows searching for different types of spatial interactions (e.g. direct and indirect cellular interaction) between objects by taking field shape into account (e.g. thin vs. broad). Third, by describing spatially distributed groups of objects as summation field, it gives cluster definition that relies rather on the bare object distance than on the modelled spatial cellular interaction.
