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Here, we introduce an amplified three-dimensional (3D) histological electrophoresis system that enables high-resolution analysis on high-throughput microwell arrays and ensures accuracy through a diffusion-analysis-supported amplification strategy. By combining targeted labeling and electrophoretic separation, the system spatially maps tumor-specific proteins in tissue sections, reconstructing malignant tissue architecture without conventional morphologic assessment. In preclinical mouse models, it accurately delineated tumor contours, characterized necrotic regions, and comprehensively detected metastatic lymph nodes. In clinical specimens, it effectively distinguished cancerous versus noncancerous regions and identified intratumoral heterogeneity. This high-throughput amplified 3D histological electrophoresis system provides a rapid and fluorescence-based diagnostic approach that complements conventional histopathological diagnosis.<\/jats:p>","DOI":"10.1021\/acsnano.6c08434","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T10:34:14Z","timestamp":1788431654000},"page":"24867-24880","source":"Crossref","is-referenced-by-count":0,"title":["Amplified\nThree-Dimensional Histological Electrophoresis\nSystem for High-Resolution Fluorescence Diagnosis of Tiny Biopsies"],"prefix":"10.1021","volume":"20","author":[{"given":"Yin","family":"Li","sequence":"first","affiliation":[{"name":"The First Hospital of Jilin University , , ,","place":["Changchun, P. R. China, 130021"]},{"name":"Jilin University , , ,","place":["Changchun, P.R. 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