Multiplex protein-specific microscopy with ultraviolet surface excitation
Multiplex protein-specific microscopy with ultraviolet surface excitation
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DOI：
10.1364/boe.11.000099
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发表时间：
2020-01-01
影响因子：
3.4
通讯作者：
Mayerich, David
中科院分区：
文献类型：
--
作者：
Guo, Jiaming;Artur, Camille;Mayerich, David
免疫组织化学技术，如免疫荧光（IF）染色，可以对组织样本中的局部蛋白质表达进行显微成像。IF的分子谱分析对于理解发病机制至关重要，并且经常涉及复杂的诊断。最近的一项创新，被称为紫外表面激发显微镜（MUSE），使用深紫外（约280 nm）照明来激发组织表面的标签，提供等效的图像，而无需固定，嵌入和切片。然而，MUSE尚未集成到传统的中频管道中。这限制了它在依赖蛋白质特异性标记的更复杂诊断中的应用。本文旨在利用量子点纳米颗粒扩大MUSE在多重免疫组织化学中的适用性。我们证明了量子点标记在石蜡包埋和完整组织上用于蛋白质特异性MUSE成像的优势，显著扩展了MUSE在蛋白质特异性应用中的适用性。此外，随着三维紫外荧光显微镜的最新创新，这为利用紫外激发量子点进行三维中频成像打开了大门。(C) 2019年美国光学学会根据OSA开放获取出版协议的条款
Immunohistochemical techniques, such as immunofluorescence (IF) staining, enable microscopic imaging of local protein expression within tissue samples. Molecular profiling enabled by IF is critical to understanding pathogenesis and is often involved in complex diagnoses. A recent innovation, known as microscopy with ultraviolet surface excitation (MUSE), uses deep ultraviolet (approximate to 280 nm) illumination to excite labels at the tissue surface, providing equivalent images without fixation, embedding, and sectioning. However, MUSE has not yet been integrated into traditional IF pipelines. This limits its application in more complex diagnoses that rely on protein-specific markers. This paper aims to broaden the applicability of MUSE to multiplex immunohistochemistry using quantum dot nanoparticles. We demonstrate the advantages of quantum dot labels for protein-specific MUSE imaging on both paraffin-embedded and intact tissue, significantly expanding MUSE applicability to protein-specific applications. Furthermore, with recent innovations in three-dimensional ultraviolet fluorescence microscopy, this opens the door to three-dimensional IF imaging with quantum dots using ultraviolet excitation. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement