title: Imaging single synaptic vesicles in Mammalian central synapses with quantum dots.
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.
authors.affiliation: Department of Pharmacology, Vanderbilt University, Nashville, TN, USA.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000818:Animals; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D011726:Pyridinium Compounds / Q000737:chemistry; D045663:Quantum Dots; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D013050:Spectrometry, Fluorescence / Q000379:methods; D013194:Staining and Labeling; D013569:Synapses / Q000378:metabolism; D009435:Synaptic Transmission; D013572:Synaptic Vesicles / Q000378:metabolism*; D046509:Tissue Culture Techniques
chemical_list: C092350:FM 4-64; D005456:Fluorescent Dyes; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds