Guo Jiaming, Artur Camille, Womack Tasha, Eriksen Jason L, Mayerich David
University of Houston, Department of Electrical and Computer Engineering, Houston, TX 77004, USA.
These authors contributed equally to this work.
Biomed Opt Express. 2019 Dec 6;11(1):99-108. doi: 10.1364/BOE.11.000099. eCollection 2020 Jan 1.
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 (MUSE), uses deep ultraviolet (≈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.
免疫组织化学技术，如免疫荧光（IF）染色，能够对组织样本中的局部蛋白质表达进行显微成像。IF实现的分子谱分析对于理解发病机制至关重要，并且常常涉及复杂的诊断。一项名为（MUSE）的最新创新技术，利用深紫外光（约280纳米）照射来激发组织表面的标记物，无需固定、包埋和切片即可提供等效图像。然而，MUSE尚未被整合到传统的IF流程中。这限制了其在依赖蛋白质特异性标记物的更复杂诊断中的应用。本文旨在通过使用量子点纳米颗粒拓宽MUSE在多重免疫组织化学中的适用性。我们展示了量子点标记物在石蜡包埋组织和完整组织上进行蛋白质特异性MUSE成像的优势，显著扩大了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，这为利用紫外激发进行量子点三维IF成像打开了大门。
