Highly Multiplexed Immunohistochemical MALDI-MS Imaging of Biomarkers in Tissues.
Highly Multiplexed Immunohistochemical MALDI-MS Imaging of Biomarkers in Tissues.
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DOI：
10.1021/jasms.0c00473
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发表时间：
2021-04-07
影响因子：
3.2
通讯作者：
Lim MJ
中科院分区：
文献类型：
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作者：
Yagnik G;Liu Z;Rothschild KJ;Lim MJ
免疫组织化学(IHC)结合荧光显微镜为研究人员和病理学家提供了一种重要的和广泛使用的工具来对组织标本中的多个生物标志物进行成像。然而，使用标准荧光显微镜的多重IHC通常仅限于3-5种不同的生物标志物，而高光谱或多光谱方法仅限于8种。我们报道了一种基于新型光裂解质谱标签(PC-MTS)的简易抗体标记新技术的发展，该技术基于MALDI质谱学成像(MALDI-IHC)实现高度多重IHC。这种方法大大超过了荧光法和以前的基于可切割质量标签的方法的多重性。在小鼠脑、人扁桃体和乳腺癌组织标本上证实了多达12-plex MALDI-IHC，反映了目标生物标记物的已知分子组成、解剖学和病理学。新的双标记荧光PC-MT抗体和无标记小分子质谱学成像极大地扩展了这种新方法的能力。MALDI-IHC在组织病理学、组织诊断学、治疗学和精密医学领域显示出良好的应用前景。
Immunohistochemistry (IHC) combined with fluorescence microscopy provides an important and widely used tool for researchers and pathologists to image multiple biomarkers in tissue specimens. However, multiplex IHC using standard fluorescence microscopy is generally limited to 3–5 different biomarkers, with hyperspectral or multispectral methods limited to 8. We report the development of a new technology based on novel photocleavable mass-tags (PC-MTs) for facile antibody labeling, which enables highly multiplexed IHC based on MALDI mass spectrometric imaging (MALDI-IHC). This approach significantly exceeds the multiplexity of both fluorescence- and previous cleavable mass-tag-based methods. Up to 12-plex MALDI-IHC was demonstrated on mouse brain, human tonsil, and breast cancer tissues specimens, reflecting the known molecular composition, anatomy, and pathology of the targeted biomarkers. Novel dual-labeled fluorescent PC-MT antibodies and label-free small-molecule mass spectrometric imaging greatly extend the capability of this new approach. MALDI-IHC shows promise for use in the fields of tissue pathology, tissue diagnostics, therapeutics, and precision medicine.
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