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TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis
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JoVE Journal Neurowissenschaften
TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis

TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis

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13:47 min

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January 29, 2015

DOI:

10.3791/52267-v

DOI:
10.3791/52267-v

•

13:47 min
January 29, 2015

•

6 Views
, , , ,
1Department of Pharmacological and Biomolecular Sciences,Università degli Studi di Milano, 2San Raffaele Scientific Institute and Vita-Salute University, 3CEND Center of Excellence in Neurodegenerative Diseases,Università degli Studi di Milano

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This paper provides a method for investigating neurotransmitter vesicle dynamics in neuroblastoma cells, using a synaptobrevin2-pHluorin construct and Total Internal Reflection Fluorescence Microscopy. The strategy developed for image processing and data analysis is also reported.

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TIRFMPH-sensitive GFP-probesNeurotransmitter Vesicle DynamicsSH-SY5Y Neuroblastoma CellsCell ImagingData AnalysisSynaptic VesiclesExo-endocytosisTotal Internal Reflection Fluorescence MicroscopySynapto-pHluorinMembrane DepolarizationNeurotransmitter ReleaseFusion Events

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Daniele, F., Di Cairano, E. S., Moretti, S., Piccoli, G., Perego, C. TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis. J. Vis. Exp. (95), e52267, doi:10.3791/52267 (2015).

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