代漂白扩展了核心设施的染色自动化
Anna E Tseng1, Aoife O'Connell2, Aaron Benhamou3
1National Emerging Infectious Diseases Laboratories (NEIDL), Boston University; Department of Pathology and Laboratory Medicine, Boston University Chobanian and Avedisian School of Medicine.
Journal of visualized experiments : JoVE
|August 25, 2025
概括
自动白扩展多重性 (IBEX) 增强了组织中的蛋白质分析. 这种胺信号放大技术提高了使用自动体的核心设施的吞吐量和可重复性.
科学领域:
- 生物医学成像
- 分子病理学
- 免疫组织化学
背景情况:
- 代漂白扩展多重性 (IBEX) 是一种基于胺信号放大 (TSA) 的技术,用于单个组织样本中的多重蛋白质成像.
- 目前的IBEX实施通常需要手动步骤,限制吞吐量和可重复性.
- 核心设施中的IBEX自动化可以显著提高效率和标准化.
研究的目的:
- 证明在常见的自动支架平台上成功应用自动化IBEX.
- 为固定在甲中的嵌入式 (FFPE) 组织设计IBEX面板提供优化策略.
- 引导核心病理设施将自动化IBEX集成到他们的工作流程中.
主要方法:
- 在两个独立的自动化平台上实施IBEX.
- 开发用于测试面板设计的抗体-抗原检索敏感性的方法.
- 使用全片扫描仪进行成像和细胞表型软件进行分析.
主要成果:
- 在两个自动化平台上成功应用了自动化IBEX.
- 在FFPE样本中基于TSA的面板设计的详细考虑,包括抗体-抗原检索敏感性测试.
- 使用商业整片扫描仪和细胞表型软件进行下游成像和分析的演示.
结论:
- 自动化IBEX具有多功能,可以轻松集成到现有的核心设施自动化培训平台中.
- 该研究提供了优化IBEX实施和面板设计的实用指导和方法.
- IBEX的自动化为多重蛋白质分析提供了更高的吞吐量,可重复性和效率.
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