Related Experiment Video
Updated: May 23, 2025
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Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes
Published on: May 8, 2016
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Standardized Flow Cytometry Method for Absolute Counting of Intraepithelial Lymphocytes in the Intestinal Mucosa
Corentin Joulain1, Stéphanie Bessoles1, Andrada S Chiron1,2
1Université Paris Cité, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, Paris, France.
Bio-Protocol
|May 21, 2025
Summary
This study introduces a flow cytometry method to accurately count intraepithelial lymphocytes (IELs) and intestinal epithelial cells (IECs), improving disease research. This technique enhances the reliability of IEL counts for digestive disease studies.
Area of Science:
- Immunology
- Gastroenterology
- Cell Biology
Background:
- Intraepithelial lymphocytes (IELs) are crucial for intestinal immunity and tumor surveillance.
- Anomalies in IEL counts are linked to various digestive diseases.
- Current IEL counting methods (histology, microscopy) are prone to bias and discrepancies.
Purpose of the Study:
- To develop and validate a flow cytometry assay for precise absolute quantification of IELs and intestinal epithelial cells (IECs).
- To establish a reliable method for determining the IEL/IEC ratio.
- To overcome limitations of conventional counting techniques and improve reproducibility in IEL research.
Main Methods:
- Combined a standard epithelial isolation technique with BD TruCount™ bead-based absolute counting.
- Utilized flow cytometry to quantify specific IEL (CD45+ CD326/EpCAM- CD103+CD3+) and IEC (CD45- CD326/EpCAM+) populations.
- Applied the method to a C57BL/6 mouse model.
Main Results:
- Successfully quantified absolute IEL counts and the IEL/IEC ratio using flow cytometry.
- Demonstrated the assay's ability to provide reproducible and standardized cell counts.
- Established a quantitative method for IEL and IEC assessment.
Conclusions:
- Flow cytometry with absolute counting beads offers a standardized and reproducible method for determining IEL and IEC counts and ratios.
- This assay addresses the limitations of traditional methods, enabling more reliable comparisons across studies.
- The developed technique is valuable for research in digestive diseases and mucosal immunology.
