Abstract
Immunocytochemistry and histochemistry are two most valuable immunochemistry techniques routinely used in biological laboratories. These techniques rely on the use of antibodies to label epitopes of interest in cells. At present, there is a wide range of commercially available organic dyes for labeling antibodies. However, limited extinction coefficients of organic dyes often make it difficult to achieve the optimal exposure and therefore fluorescence detection limit. Quantum dots (QDs) are fluorescent semiconductor nanocrystals which are advantageous over the organic fluorescent dyes in many aspects, principally their long-term luminescence stability, high brightness, and multicolor detection. Here, we describe the use of QDs for immunocytochemistry applications. We used three different antibodies—anti-β-tubulin monoclonal antibody for visualizing the microtubule network, the GM130 antibody for staining the Golgi complex, and the EEA1 antibody for detecting the endosomal system. We use the anti-mouse IgG antibody directly conjugated to QD655 quantum dots or anti-mouse IgG conjugated to biotin for tertiary detection with streptavidin-conjugated QD655.
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References
Dodson A (2002) Review: modern methods for diagnostic immunocytochemistry. Curr Diagn Pathol 8:113–122
Resch-Genger U, Grabolle M, Cavaliere-Jaricot S et al (2008) Quantum dots versus organic dyes as fluorescent labels. Nat Methods 5:763–775
Mazumder S, Dey R, Mitra MK et al (2009) Review: biofunctionalized quantum dots in biology and medicine. J Nanomater. doi:10.1155/2009/815734
Chan WC, Maxwell DJ, Gao X et al (2002) Luminescent quantum dots for multiplexed biological detection and imaging. Curr Opin Biotechnol 13:40–46
Tiwary DK, Tanaka S-I, Inouye Y et al (2009) Synthesis and characterization of anti-HER2 antibody conjugated CdSe/CdZnS quantum dots for fluorescence imaging of breast cancer cells. Sensors 9:9332–9354
Shi C, Zhou G, Zhu Y et al (2008) Quantum dots-based multiplexed immunohistochemistry of protein expression in human prostate cancer cells. Eur J Histochem 52:127–134
Sweeney E, Ward TH, Gray N et al (2008) Quantitative multiplexed quantum dot immunohistochemistry. Biochem Bioph Res Co 374:181–186
Montón H, Nogues C, Rossinyol E et al (2009) QDs versus Alexa: reality of promising tools for immunocytochemistry. J Nanobiotechnolgy. doi:10.1186/1477-3155-7-4
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Montón, H., Roldán, M., Merkoçi, A., Rossinyol, E., Castell, O., Nogués, C. (2012). The Use of Quantum Dots for Immunochemistry Applications. In: Soloviev, M. (eds) Nanoparticles in Biology and Medicine. Methods in Molecular Biology, vol 906. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-953-2_13
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DOI: https://doi.org/10.1007/978-1-61779-953-2_13
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-952-5
Online ISBN: 978-1-61779-953-2
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