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Super-multiplexed fluorescence nanoscopy for imaging-based proteomics

Super-multiplexed fluorescence nanoscopy for imaging-based proteomics
用于基于成像的蛋白质组学的超级多重荧光纳米镜
批准号:
10388887
负责人:
Kyu Young Han
金额:
$20.0万
依托单位国家:
美国
项目类别:
财政年份:
2020
资助国家:
美国
项目状态:
未结题
起止时间:
2020-09-15 至 2025-08-31

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中文摘要
翻译
项目摘要 原位免疫荧光成像是一种有效的方法来研究肿瘤的位置,表达水平和 细胞和组织中的蛋白质结构。特别是,多重成像揭示了相互作用网络 这使我们能够了解许多疾病的潜在机制。但却 由于其极其耗时, 工艺复杂、成本高、信号放大不足。共焦显微镜有限的空间分辨率 显微镜通常不能揭示复杂的空间组织和确定蛋白质的定位。这里 我们提出了一种超多路复用免疫荧光纳米显微镜, 24小时内不同的蛋白质。我们将使用DNA条形码化的二级纳米抗体, 它们是单价的、开源的,专为定量标记而设计。反复导入和清洗 荧光DNA成像仪将产生高度多路复用的图像。此外,我们将开发 前所未有的快速受激发射耗尽(STED)显微镜,采用平行线阵列 甜甜圈梁。它将具有大的成像面积和出色的光学切片能力。光子 重新分配,高光谱成像和深度学习将进一步促进基于快速超分辨率的 蛋白质谱分析我们新的生物化学和光学工具将在不同的生物医学领域发挥关键作用 包括大脑蛋白质组学和癌症分析。
英文摘要
PROJECT SUMMARY In situ immunofluorescence imaging is a powerful method to study the locations, expression levels and structures of proteins in cells and tissues. In particular, multiplexed imaging reveals the interaction networks of proteins, which allows us to understand the underlying mechanisms of many diseases. However, it has been challenging to perform multiplexed immunofluorescence imaging due to its extremely time-consuming process, high cost and lack of signal amplification. The limited spatial resolution achievable with confocal microscopy often fails to reveal complex spatial organization and to determine localizations of proteins. Here we propose super-multiplexed immunofluorescence nanoscopy that is capable of imaging more than twenty different proteins in 24 hours with nanoscale resolution. We will employ DNA-barcoded secondary nanobodies that are monovalent, open-source and designed for quantitative labeling. Repeated introduction and washing of fluorescent DNA imagers will generate highly multiplexed images. Moreover, we will develop unprecedentedly fast stimulated emission depletion (STED) microscopy that employs a parallelized line array of doughnut beams. It will feature a large imaging area and excellent optical sectioning capability. Photon reassignment, hyperspectral imaging and deep-learning will further facilitate rapid super-resolution-based protein profiling. Our new biochemical and optical tools will play crucial roles in diverse biomedical areas including brain proteomics and cancer profiling.
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Super-multiplexed fluorescence nanoscopy for imaging-based proteomics
  • 批准号:
    10028050
  • 项目类别:
  • 资助金额:
    $26.76万
  • 财政年份:
    2020
  • 负责人:
    Kyu Young Han
  • 依托单位:
Super-multiplexed fluorescence nanoscopy for imaging-based proteomics
  • 批准号:
    10471273
  • 项目类别:
  • 资助金额:
    $34.58万
  • 财政年份:
    2020
  • 负责人:
    Kyu Young Han
  • 依托单位:
Super-multiplexed fluorescence nanoscopy for imaging-based proteomics
  • 批准号:
    10261562
  • 项目类别:
  • 资助金额:
    $33.89万
  • 财政年份:
    2020
  • 负责人:
    Kyu Young Han
  • 依托单位:
海外基金