Abstract
This protocol describes a sensitive and rigorous method to monitor the movement and turnover of single synaptic vesicles in live presynaptic terminals of mammalian central nervous system. This technique makes use of Photoluminescent semiconductor nanocrystals, quantum dots (Qdots), by their nanometer size, superior photoproperties, and pH-sensitivity. In comparison with previous fluorescent probes like styryl dyes and pH-sensitive fluorescent proteins, Qdots offer strict loading ratio, multi-modality detection, single vesicle precision, and most importantly distinctive signals for different modes of vesicle fusion. Qdots are spectrally compatible with existing fluorescent probes for synaptic vesicles and thus allow multichannel imaging. With easy modification, this technique can be applied to other types of synapses and cells.
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Acknowledgments
I thank my postdoctoral mentor, Dr. R. W. Tsien, for giving me the opportunity to develop this Qdot-based vesicle labeling method in his lab. I also thank Invitrogen for providing the documentation of Qdots’ properties. This work was supported by grants from NIDA and AFAR to Q.Z.
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Zhang, Q. (2013). Imaging Single Synaptic Vesicles in Mammalian Central Synapses with Quantum Dots. In: Banghart, M. (eds) Chemical Neurobiology. Methods in Molecular Biology, vol 995. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-345-9_13
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DOI: https://doi.org/10.1007/978-1-62703-345-9_13
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