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Updated: Jun 10, 2025
Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Published on: January 9, 2019
Multiplexed Immunophenotyping of Lymphoma Tissue Samples
Christina Schniederjohann1,2,3,4,5, Peter-Martin Bruch1,2,3,4, Sascha Dietrich6,7,8,9
1Department of Hematology, Oncology and Clinical Immunology, University Hospital Düsseldorf, Düsseldorf, Germany.
Insights
High-plex imaging, like CO-Detection by IndEXing (CODEX), reveals cellular details in lymphoma tissue. This technique enhances understanding of tumor and microenvironment interactions for better diagnostics and therapies.
Area of Science:
- Biomedical Imaging
- Immunohistochemistry
- Molecular Pathology
Background:
- High-plex imaging allows detailed analysis of cellular phenotypes and interactions in tissue biopsies.
- Lymphoma research benefits from single-cell resolution to understand tumor and microenvironmental components.
- Investigating biological mechanisms and biomarkers requires advanced imaging techniques.
Purpose of the Study:
- To provide an overview of key steps in a CO-Detection by IndEXing (CODEX) project.
- To share a CODEX protocol and experience with FFPE tissue samples for lymphoma research.
- To encourage wider adoption of CODEX for improved lymphoma diagnostics and therapy.
Main Methods:
- Antibody panel design for high-plex imaging.
- Cohort selection for tissue sample analysis.
- Staining, imaging, and data analysis procedures for CODEX.
Main Results:
- Detailed characterization of cellular phenotypes and interactions in lymphoma.
- Insights into spatial dynamics of tumor and microenvironmental cells.
- Demonstration of CODEX applicability on FFPE tissue samples.
Conclusions:
- CODEX is a powerful technique for lymphoma research, offering single-cell resolution.
- The shared protocol facilitates wider use and deeper understanding of lymphoma biology.
- Enhanced insight into cellular composition and spatial dynamics can improve diagnostics and therapy.
Abstract:
High-plex imaging techniques enable the detection and quantification of a multitude of markers in tissue biopsies at single-cell or near-single-cell resolution. In lymphoma, this can facilitate the detection and characterization of cellular phenotypes and interactions, describing both tumor and microenvironmental cells. In combination with other techniques, high-plex imaging allows the investigation of biological mechanisms and clinically relevant biomarkers. CO-Detection by IndEXing (CODEX), one of such techniques, is based on antibodies labeled with unique DNA oligonucleotides that can be visualized by complementary reporter oligonucleotides coupled to a fluorophore. Here, we provide an overview of the key steps of a CODEX-based project, including (1) antibody panel design, (2) cohort selection, (3) staining and imaging, (4) data analysis. By sharing our CODEX protocol and our experience with FFPE tissue samples, we aim to encourage wider use of this powerful technique in lymphoma research and improve insight into cellular composition and spatial dynamics for improved diagnostics and therapy.
