Abstract
Single-particle tracking (SPT) using fluorescent quantum dots (QDs) provides high-resolution spatial–temporal information on receptor dynamics that cannot be obtained through traditional biochemical techniques. In particular, the high brightness and photostability of QDs make them ideal probes for SPT on living cells. We have shown that QD-labeled IgE can be used to characterize the dynamics of the high-affinity IgE Receptor. Here, we describe protocols for (1) coupling QDs to IgE, (2) tracking individual QD-bound receptors, and (3) analyzing one- and two-color tracking data.
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Acknowledgments
This work was supported by NIH P50GM085273, ACS-IRG #192, and the Human Frontier Science Program.
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Lidke, D.S., Low-Nam, S.T., Cutler, P.J., Lidke, K.A. (2011). Determining FcεRI Diffusional Dynamics via Single Quantum Dot Tracking. In: Rast, J., Booth, J. (eds) Immune Receptors. Methods in Molecular Biology, vol 748. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-139-0_8
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DOI: https://doi.org/10.1007/978-1-61779-139-0_8
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