Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry.
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.
Xing
Yun
Y
Department of Biomedical Engineering, Emory University and Georgia Institute of Technology, 101 Woodruff Circle Suite 1001, Atlanta, Georgia 30322, USA.
Chaudry
Qaiser
Q
Shen
Christopher
C
Kong
Koon Yin
KY
Zhau
Haiyen E
HE
Chung
Leland W
LW
Petros
John A
JA
O'Regan
Ruth M
RM
Yezhelyev
Maksym V
MV
Simons
Jonathan W
JW
Wang
May D
MD
Nie
Shuming
S
D000906:Antibodies / Q000378:metabolism; D015415:Biomarkers / Q000032:analysis; D007150:Immunohistochemistry / Q000295:instrumentation* / Q000379:methods*; D045663:Quantum Dots; D046509:Tissue Culture Techniques / Q000379:methods
D000906:Antibodies; D015415:Biomarkers