Because of their unique fluorescence properties, quantum dots (QDs) represent a promising new technology in the realm of multicolor flow cytometry. Although commercial reagents and applications for the technology are still in the early phases of their development, the strategies and considerations necessary for successful use are becoming known. This article discusses the value of QDs in multicolor flow cytometry, introduces strategies to successfully incorporate QDs into routine use, and highlights emerging applications of the technology. WIREs Nanomed Nanobiotechnol 2010 2 334–348
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"Every separate combination of antibodies that is to be stored as a cocktail requires a shelf-life validation. There may also be certain reagents that are not suitable for storage in a cocktail; for instance, the non-organic q-dots are sensitive to changes in their storage media, and may form aggregates and/ or bind plastic at low volumes (Jaiswal & Simon, 2004; Biancotto et al, 2011; Chattopadhyay et al, 2012). The current collective experience of this writing group is that home-made cocktails containing eight or more antibodies conjugated to organic fluorochromes, including tandem fluorochromes, work well. "
"Still, users who rely on UV-excited probes (like DAPI and Hoechst) should note that QDs are compatible with their systems (Telford WG, 2004). Wheremultiplexed analysis of QDs is important, UV or violet excitation systems can be coupled to as many as eight photomultiplier tubes, allowing simultaneous measurement of QD545, QD565, QD585, QD605, QD655, QD705, QD800, and/or a violetexcitable organic fluorochrome (Chattopadhyay et al, 2010). To detect QD signals, we employ a filter strategy that first selects light sharply with a dichroic mirror, allowing only light above a certain wavelength to pass (long-pass filter). "
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