ReviewJournal of translational medicine2026
Spotlight on challenges and novel methods in highly multiplexed tissue imaging-based spatial proteomics.
Review in Journal of translational medicine, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Authors and funding
2 authors.
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Abstract
backgroundTissue microenvironment heterogeneity, including cellular composition, functional states, and intercellular interactions, shapes complex biological processes and disease mechanisms. Recent progress in highly multiplexed tissue imaging (HMTI) has enabled single-cell-resolved spatial proteomics, supporting detailed interrogation of dynamic tissue organization across contexts such as cancer, embryonic development, and neuroscience. However, the rapidly expanding HMTI landscape also creates practical challenges in platform selection, computational analysis, and cross-study reproducibility. MAIN BODY: HMTI platforms leverage antibody labeling strategies, including DNA oligonucleotide-tagged, fluorophore-tagged, and metal-tagged reagents, to capture highly multiplexed molecular readouts (often tens to > 100 markers) within the same specimen. These approaches generate rich spatially indexed datasets that can reveal cell neighborhoods, spatial programs, and interaction patterns. In this review, we provide a comprehensive comparison of representative HMTI platforms and highlight major analytical bottlenecks in HMTI data analysis, including preprocessing and normalization, denoising, spillover correction, cell segmentation, cell-type annotation, and tissue microarchitecture analysis, with particular emphasis on method limitations, scalability, and performance under challenging conditions. In addition, we discuss tissue- and disease-specific parameter selection, pixel-level analysis, integration with spatial multi-omics, emerging three-dimensional spatial proteomics, and current barriers to clinical translation.
conclusionsHMTI-driven spatial proteomics provides a powerful route to quantify tissue microenvironment heterogeneity at single-cell resolution. Future progress will depend on improved standardized and scalable computational pipelines, robust benchmarking datasets, and more reliable multimodal and three-dimensional analytical frameworks. These advances are expected to enhance reproducibility and accelerate the translation of HMTI toward clinically actionable applications in precision medicine.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.