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Highly Multiplexed Single‐Cell Protein Profiling with Large‐Scale Convertible DNA‐Antibody Barcoded Arrays

机译:具有大规模可转换DNA抗体条形码阵列的高度多路复用单细胞蛋白质分析

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摘要

Highly multiplexed detection of proteins secreted by single cells is always challenging. Herein, a multiplexed in situ tagging technique based on single‐stranded DNA encoded microbead arrays and multicolor successive imaging for assaying single‐cell secreted proteins with high throughput and high sensitivity is presented. This technology is demonstrated to be capable of increasing the multiplexity exponentially. Upon integration with polydimethylsiloxane microwells, this platform is applied to detect ten immune effector proteins from differentiated single macrophages stimulated with lipopolysaccharide. Significant heterogeneity is observed when the derived human primary macrophages are analyzed. This versatile technology is expected to open new opportunities in systems biology, immune regulation studies, signaling analysis, and molecular diagnostics.
机译:高度多重检测单个细胞分泌的蛋白质始终具有挑战性。本文提出了一种基于单链DNA编码的微珠阵列和多色连续成像的多重原位标记技术,以高通量和高灵敏度测定单细胞分泌的蛋白质。事实证明,该技术能够以指数方式增加复用性。与聚二甲基硅氧烷微孔整合后,该平台可用于从脂多糖刺激的分化的单个巨噬细胞中检测十种免疫效应蛋白。分析派生的人类原代巨噬细胞时,观察到明显的异质性。这种多功能技术有望为系统生物学,免疫调节研究,信号分析和分子诊断等领域带来新的机遇。

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