Xing Yun, Chaudry Qaiser, Shen Christopher, Kong Koon Yin, Zhau Haiyen E, Chung Leland W, Petros John A, O'Regan Ruth M, Yezhelyev Maksym V, Simons Jonathan W, Wang May D, Nie Shuming
Department of Biomedical Engineering, Emory University and Georgia Institute of Technology, 101 Woodruff Circle Suite 1001, Atlanta, Georgia 30322, USA.
Nat Protoc. 2007;2(5):1152-65. doi: 10.1038/nprot.2007.107.
Bioconjugated quantum dots (QDs) provide a new class of biological labels for evaluating biomolecular signatures (biomarkers) on intact cells and tissue specimens. In particular, the use of multicolor QD probes in immunohistochemistry is considered one of the most important and clinically relevant applications. At present, however, clinical applications of QD-based immunohistochemistry have achieved only limited success. A major bottleneck is the lack of robust protocols to define the key parameters and steps. Here, we describe our recent experience, preliminary results and detailed protocols for QD-antibody conjugation, tissue specimen preparation, multicolor QD staining, image processing and biomarker quantification. The results demonstrate that bioconjugated QDs can be used for multiplexed profiling of molecular biomarkers, and ultimately for correlation with disease progression and response to therapy. In general, QD bioconjugation is completed within 1 day, and multiplexed molecular profiling takes 1-3 days depending on the number of biomarkers and QD probes used.
生物共轭量子点（QDs）为评估完整细胞和组织标本上的生物分子特征（生物标志物）提供了一类新型生物标记物。特别是，多色量子点探针在免疫组织化学中的应用被认为是最重要且与临床相关的应用之一。然而目前，基于量子点的免疫组织化学的临床应用仅取得了有限的成功。一个主要瓶颈是缺乏确定关键参数和步骤的可靠方案。在此，我们描述了我们在量子点-抗体共轭、组织标本制备、多色量子点染色、图像处理和生物标志物定量方面的近期经验、初步结果和详细方案。结果表明，生物共轭量子点可用于分子生物标志物的多重分析，并最终用于与疾病进展和治疗反应的关联分析。一般来说，量子点生物共轭在1天内完成，多重分子分析根据所使用的生物标志物和量子点探针的数量需要1至3天。
