AmberGen, Inc., 313 Pleasant Street, Watertown, Massachusetts 02472, United States.
Molecular Biophysics Laboratory, Department of Physics and Photonics Center, Boston University, Boston, Massachusetts 02215, United States.
J Am Soc Mass Spectrom. 2021 Apr 7;32(4):977-988. doi: 10.1021/jasms.0c00473. Epub 2021 Feb 25.
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.
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且广泛使用的工具，用于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 通常限于 3-5 种不同的生物标志物，而超光谱或多光谱方法限于 8 种。我们报告了一种基于新型光可裂解质量标签（PC-MTs）的新技术的开发，用于简便的抗体标记，该技术可基于 MALDI 质谱成像（MALDI-IHC）实现高度多重化的 IHC。这种方法大大超过了荧光和以前的可裂解质量标记方法的多重性。在小鼠脑、人扁桃体和乳腺癌组织标本上证明了多达 12 重的 MALDI-IHC，反映了靶向生物标志物的已知分子组成、解剖结构和病理学。新型双重标记荧光 PC-MT 抗体和无标记小分子质谱成像极大地扩展了这种新方法的能力。MALDI-IHC 有望在组织病理学、组织诊断、治疗和精准医学领域得到应用。
