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Updated: Jul 6, 2025

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
ESQmodel: biologically informed evaluation of 2-D cell segmentation quality in multiplexed tissue images
Eric Lee1,2, Dongkyu Lee3, Wayne Fan4
1Department of Molecular Oncology, BC Cancer Agency, Vancouver, British Columbia V5Z1L3, Canada.
Motivation:
Single cell segmentation is critical in the processing of spatial omics data to accurately perform cell type identification and analyze spatial expression patterns. Segmentation methods often rely on semi-supervised annotation or labeled training data which are highly dependent on user expertise. To ensure the quality of segmentation, current evaluation strategies quantify accuracy by assessing cellular masks or through iterative inspection by pathologists. While these strategies each address either the statistical or biological aspects of segmentation, there lacks a unified approach to evaluating segmentation accuracy.
Results:
In this article, we present ESQmodel, a Bayesian probabilistic method to evaluate single cell segmentation using expression data. By using the extracted cellular data from segmentation and a prior belief of cellular composition as input, ESQmodel computes per cell entropy to assess segmentation quality by how consistent cellular expression profiles match with cell type expectations.
Availability And Implementation:
Source code is available on Github at: https://github.com/Roth-Lab/ESQmodel.
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