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Updated: Jan 13, 2026
Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes
Published on: May 8, 2016
A Paired Flow Cytometry-Pathology Assessment for Immune Cell Detection in Intestinal Biopsies: Proof of Principle
Alexandros Skamnelos1, Georgios S Markopoulos2,3, Lefkothea Dova2
1Department of Gastroenterology, University Hospital of Ioannina, 45110 Ioannina, Greece.
Insights
This study introduces a combined flow cytometry and pathology method for immune cell quantification in colon biopsies. The integrated approach enhances diagnostic accuracy by cross-validating results from both techniques.
Area of Science:
- Immunology
- Pathology
- Biotechnology
Background:
- Accurate immune cell quantification is crucial for research and clinical applications.
- Flow cytometry (FC) offers single-cell resolution, while pathology provides spatial context.
- Current methods analyze immune cells separately, limiting comparative analysis.
Purpose of the Study:
- To develop and validate an integrated workflow combining FC and pathology for immune cell quantification.
- To assess immune cell populations within the same colon biopsy specimen.
- To improve diagnostic accuracy through cross-validation of methodologies.
Main Methods:
- Colon biopsy samples were divided for FC and formalin-fixed, paraffin-embedded (FFPE) pathology analysis.
- FC characterized CD45+, CD3+, CD4+, and CD8+ T cells.
- Pathology utilized H&E staining for eosinophils and IHC for CD4 and CD8.
Main Results:
- A pilot study of 10 samples demonstrated high concordance between FC and pathology results.
- The integrated approach allowed for cross-validation of immune cell quantification.
- Feasibility of combining FC and pathology on limited intestinal biopsy material was shown.
Conclusions:
- An integrated workflow combining FC and pathology is feasible for immune cell quantification in colon biopsies.
- This method enables comprehensive assessment of immune cell populations.
- The approach holds potential for improved diagnostic accuracy in immunological studies.
Abstract:
Accurate quantification of immune cell subpopulations is essential for understanding immune responses in research and clinical settings. Flow cytometry (FC) is widely used for immune cell phenotyping, providing rapid and quantitative single-cell resolution. However, tissue-based pathological assessment offers additional spatial and morphological context that is often necessary for a comprehensive understanding of immune cell distribution. Traditionally, these methods are applied separately to different specimens, limiting direct comparative analysis. Here, we describe a simple combined approach to immune cell quantification that integrates both FC and pathology analysis within the same tissue specimen of colon biopsies. Tissue samples were divided into two portions: one processed into a single-cell suspension for FC-based characterization of CD45+, CD3+, CD4+, and CD8+ T cells and another formalin-fixed, paraffin-embedded (FFPE), and analyzed with hematoxylin and eosin (H&E) for eosinophils and immunohistochemistry (IHC) for CD4 and CD8. A pilot analysis of 10 samples shows high concordance of the results taken from the two methods, allowing for cross-validation between methodologies and improved diagnostic accuracy. This proof-of-principle study demonstrates the feasibility of an integrated workflow that combines FC and pathology for immune cell quantification, which provides assessment of immune cell populations from the limited material of intestinal biopsies with potential for improved diagnostic accuracy.
