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Image-Based Cell Profiling Enables Quantitative Tissue Microscopy in Gastroenterology.

John W Wills1, Jack Robertson1, Huw D Summers2

  • 1Biominerals Research, Cambridge University Department of Veterinary Medicine, School of Biological Sciences, Cambridge, UK.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|May 24, 2020
PubMed
Summary
This summary is machine-generated.

This study introduces a user-friendly method for quantitative tissue microscopy, enabling detailed cell analysis in diverse gastrointestinal tissues across species. The approach enhances data reproducibility and offers new insights into gastroenterology research.

Keywords:
cell segmentationconfocal microscopyimmunofluorescenceintestinal tissuemachine learningprocessing tilescans in CellProfiler | Getis-Ord spatial statistics

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Area of Science:

  • * Biomedical imaging
  • * Cell biology
  • * Gastroenterology

Background:

  • * Immunofluorescence microscopy is crucial for tissue research but often yields only qualitative data.
  • * Quantifying per-cell data in complex tissues like the gastrointestinal tract is challenging due to cellular diversity and species variation.
  • * Existing methods struggle to provide location-specific, quantitative cellular information.

Purpose of the Study:

  • * To develop an open, user-friendly methodology for quantitative tissue microscopy applicable to diverse gastrointestinal tissues, species, and preparation types.
  • * To enable "flow cytometry-type" analyses with preserved spatial data for per-cell quantification.
  • * To address the limitations of qualitative microscopy and improve data reproducibility in tissue-based research.

Main Methods:

  • * Optimized antibody-independent cell labeling compatible with various tissue preparations and species.
  • * Extracted per-cell data from confocal micrographs using semantic machine learning for dense tissues.
  • * Performed flow cytometry-type analyses, visualization, and statistical mapping of cell locations, interactions, and microenvironments.

Main Results:

  • * Successfully quantified rare events, such as *Escherichia coli* passage in human small bowel tissue.
  • * Differentiated and quantified intraepithelial lymphocyte populations in rat jejunum relative to enterocytes.
  • * Mapped mononuclear phagocyte-T cell interactions and spatial congregations in mouse Peyer's patches.

Conclusions:

  • * Accessible quantitative tissue microscopy offers novel insights into gastroenterological research questions.
  • * The developed methodology enhances data reproducibility and overcomes limitations of qualitative imaging and antibody reliance.
  • * This approach facilitates detailed analysis of cell location, interactions, and microenvironments within complex tissues.