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Part of the series Springer Protocols Handbooks pp 1-12

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Quantum Dot Toolbox in Membrane Neurotransmitter Transporter Research

  • Lucas B. ThalAffiliated withDepartment of Chemistry, Vanderbilt University
  • , Danielle M. BaileyAffiliated withDepartment of Chemistry, Vanderbilt UniversityDepartment of Pharmacology, Vanderbilt UniversityDepartment of Interdisciplinary Materials Science, Vanderbilt University
  • , Oleg KovtunAffiliated withDepartment of Chemistry, Vanderbilt University
  • , Sandra J. RosenthalAffiliated withDepartment of Chemistry, Vanderbilt UniversityDepartment of Pharmacology, Vanderbilt UniversityDepartment of Interdisciplinary Materials Science, Vanderbilt UniversityDepartment of Chemical and Biomolecular Engineering, Vanderbilt UniversityDepartment of Physics and Astronomy, Vanderbilt UniversityVanderbilt Institute of Nanoscale Science and Engineering, Vanderbilt UniversityMaterials Science and Technology Division, Oak Ridge National Laboratory Email author 

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Abstract

Quantum dot-based fluorescence techniques enable multi-scale molecular profiling ranging from real-time single molecule dynamics to expression trends in million-cell populations. In comparison to currently available probes, quantum dots are particularly well suited for such studies by virtue of their unique photophysical properties. We discuss in this chapter methodological components of what makes up the “Quantum Dot Toolbox” in neurotransmitter transporter studies along with specific work our group has published. First, we describe ensemble analysis of subcellular transporter localization and provide visualization of transporter residence in distinct cellular surface features. Second, we provide discussion on high content analysis of changes in transporter surface levels and give insight into the advantages of using quantum dot probes in flow cytometry. Third, we review the fundamental principles of subdiffraction-limit fluorescence microscopy and single molecule analysis of transporter surface dynamics. Included in this chapter are three protocols with experimental considerations specific to each technical section.

Keywords:

Flow cytometry Neurotransmitter transporters Quantum dot-based fluorescence Quantum dots Subcellular transporter localization