Protocol abstract


Nature Protocols 2, 1152 - 1165 (2007)
Published online: 3 May 2007 | doi:10.1038/nprot.2007.107

Subject Categories: Imaging | Nanotechnology | Chemical modification

Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry

Yun Xing1, Qaiser Chaudry2, Christopher Shen1, Koon Yin Kong2, Haiyen E Zhau3, Leland W Chung3, John A Petros3,4, Ruth M O'Regan5, Maksym V Yezhelyev5, Jonathan W Simons1,5, May D Wang1,2,5 & Shuming Nie1,5


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.

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  1. Department of Biomedical Engineering, Emory University and Georgia Institute of Technology, 101 Woodruff Circle Suite 1001, Atlanta, Georgia 30322, USA.
  2. Department of Electrical and Computer Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
  3. Department of Urology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
  4. Veteran Affairs Medical Center, Atlanta, Georgia, 30333, USA.
  5. Department of Hematology and Oncology and the Winship Cancer Institute, Emory University School of Medicine, Atlanta, Georgia 30322, USA.

Correspondence to: May D Wang1,2,5 e-mail: maywang@bme.gatech.edu

Correspondence to: Shuming Nie1,5 e-mail: snie@emory.edu

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