caa a22 4500 445303085 CHVBK 20180317142604.0 cr unu---uuuuu 170323e20100101xx s 000 0 eng 10.1007/s12274-010-1009-1 doi (NATIONALLICENCE)springer-10.1007/s12274-010-1009-1 Comparison and optimization of multiplexed quantum dot-based immunohistofluorescence [Elektronische Daten] [Donghai Huang, Xianghong Peng, Ling Su, Dongsheng Wang, Fadlo Khuri, Dong Shin, Zhuo Chen] Nanoparticle quantum dots (QDs) are ideal materials for multiplexed biomarker detection, localization, and quantification. Both direct and indirect methods are available for QD-based immunohistofluorescence (QD-IHF) staining; the direct method, however, has been considered laborious and costly. In this study, we optimized and compared the indirect QD-IHF single staining procedure using QD-secondary antibody conjugates and QD-streptavidin conjugates. Problems associated with sequential multiplex staining were identified quantitatively. A method using a QD cocktail solution was developed allowing simultaneous staining with three antibodies against E-cadherin, epidermal growth factor receptor and β-catenin in formalin-fixed and paraffin-embedded (FFPE) tissues. The expression of each biomarker was quantified by using the cocktail and the sequential methods. Comparison of the two methods demonstrated that the cocktail method provided more consistent and stable QD signals for each multiplexed biomarker than the sequential method, and provides a convenient tool for multiplexing biomarkers in both research and clinical applications. Tsinghua University Press and Springer Berlin Heidelberg, 2010 Quantum dots nationallicence immunohistofluorescence nationallicence nanocrystals nationallicence spectral imaging nationallicence Huang Donghai Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut Peng Xianghong Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut Su Ling Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut Wang Dongsheng Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut Khuri Fadlo Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut Shin Dong Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut Chen Zhuo Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut Nano Research Tsinghua Press 3/1(2010-01-01), 61-68 1998-0124 3:1<61 2010 3 12274 https://doi.org/10.1007/s12274-010-1009-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12274-010-1009-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Huang Donghai Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut NATIONALLICENCE 700 1- Peng Xianghong Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut NATIONALLICENCE 700 1- Su Ling Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut NATIONALLICENCE 700 1- Wang Dongsheng Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut NATIONALLICENCE 700 1- Khuri Fadlo Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut NATIONALLICENCE 700 1- Shin Dong Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut NATIONALLICENCE 700 1- Chen Zhuo Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, 30322, Atlanta, Georgia, USA aut NATIONALLICENCE 773 0- Nano Research Tsinghua Press 3/1(2010-01-01), 61-68 1998-0124 3:1<61 2010 3 12274 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer