Huang, Donghai and Peng, Xianghong and Su, Ling and Wang, Dongsheng and Khuri, Fadlo R. and Shin, Dong M. and Chen, Zhuo (Georgia) (2009) Comparison and optimization of multiplexed quantum dot-based immunohistofluorescence. Nano Research, 3 (1). pp. 61-68.
Official URL: htp://www.thenanoresearch.com/files/61.pdf
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 Î²-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.
|Subjects:||Physical Science > Nanophysics|
Physical Science > Nano objects
Material Science > Nanochemistry
Material Science > Nanostructured materials
|Divisions:||Faculty of Engineering, Science and Mathematics > School of Physics|
Faculty of Engineering, Science and Mathematics > School of Chemistry
|Deposited On:||21 May 2010 10:53|
|Last Modified:||21 May 2010 10:53|
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