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Advancing multiplexed imaging for enhanced tissue complexity analysis

Nature Methods volume 20, pages 1280–1281 (2023)Cite this article

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Multiplexed spatial immunophenotyping has advanced our understanding of tissues in the context of homeostasis and disease. Two studies now provide additional tools to overcome challenges with multiplexed imaging: one procedure amplifies the detection of low-abundance antigens by integrating SABER and IMC technologies, and the other is an X-ray-based method that enables the non-destructive multiplexed detection of antigens in tissues at scalable resolution and speed.

Multidimensional approaches that enable the analysis of various parameters at the single-cell level have transformed our understanding of biological systems. Moreover, the development of multiplex immunophenotyping approaches that preserve the spatial context of cells — such as co-detection by indexing (CODEX)6, multiplexed ion beam imaging (MIBI)7 and IMC1 — has provided unprecedented insights into the multicellular architecture of tissues. IMC achieves its multiplexing capacity by simultaneously measuring metal isotope-labeled antibodies, which are detected in a mass cytometer. Although IMC has already provided important insights into various pathologies (including the characterization of tumor microenvironments)8, a major limitation of IMC and other state-of-the-art spatial immunophenotyping methodologies is the absence of a signal amplification step, which makes it difficult to detect low-abundance antigens. These include several immunomodulatory molecules (for example, PD-1, PD-L1 and CTLA-4) that are essential for defining immune cell subsets of functional and clinical relevance. In recent years, a few methodologies that support the signal amplification of multiplex immunodetection have been developed, including tyramide signal amplification and immuno-SABER. The latter was developed in 2019 by Saka et al.4 to enable the simultaneous amplified detection of up to ten proteins using DNA-barcoded antibodies and fluorescently labeled DNA concatemers. These concatemers form branched DNA structures on the antibodies, which results in a 5- to 180-fold signal amplification (depending on the number of branching steps performed).

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Fig. 1: SABERx-IMC and MEZ-XRF are solutions for challenges in IMC.

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Acknowledgements

N.F.C.C.d.M. is funded by the European Research Council (ERC) under the European Union’s Horizon 2020 Research and Innovation Programme (grant agreement no. 852832).

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  1. Pathology, Leiden University Medical Center, Leiden, The Netherlands

    Marieke E. Ijsselsteijn & Noel F. C. C. de Miranda

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  1. Marieke E. Ijsselsteijn
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  2. Noel F. C. C. de Miranda
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Correspondence to Noel F. C. C. de Miranda.

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Ijsselsteijn, M.E., de Miranda, N.F.C.C. Advancing multiplexed imaging for enhanced tissue complexity analysis. Nat Methods 20, 1280–1281 (2023). https://doi.org/10.1038/s41592-023-01935-7

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