Department of Hematology, Oncology and Clinical Immunology, University Hospital Düsseldorf, Düsseldorf, Germany.
Center for Integrated Oncology Aachen-Bonn-Cologne-Düsseldorf (CIO ABCD), Düsseldorf, Germany.
Methods Mol Biol. 2025;2865:375-393. doi: 10.1007/978-1-0716-4188-0_16.
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.
高多重成像技术能够以单细胞或接近单细胞分辨率检测和定量组织活检中的多种标记物。在淋巴瘤中，这可以促进细胞表型和相互作用的检测和特征描述，包括肿瘤细胞和微环境细胞。与其他技术结合使用时，高多重成像可以研究生物机制和临床相关的生物标志物。CODEX 是其中一种技术，它基于带有独特 DNA 寡核苷酸的抗体，这些寡核苷酸可以通过与荧光团偶联的互补报告寡核苷酸进行可视化。在这里，我们提供了基于 CODEX 的项目的关键步骤概述，包括（1）抗体面板设计，（2）队列选择，（3）染色和成像，（4）数据分析。通过分享我们的 CODEX 方案和我们在 FFPE 组织样本方面的经验，我们旨在鼓励在淋巴瘤研究中更广泛地使用这项强大的技术，并深入了解细胞组成和空间动态，以提高诊断和治疗效果。
