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JoVE Journal Neuroscience
TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells
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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

Article DOI: 10.3791/52267-v • 13:47 min • January 29th, 2015
January 29th, 2015 •
1, 1, 1, 2, 1,3
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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Summary
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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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TIRFM PH-sensitive GFP-probes Neurotransmitter Vesicle Dynamics SH-SY5Y Neuroblastoma Cells Cell Imaging Data Analysis Synaptic Vesicles Fusion And Retrieval Exo-endocytosis Real-time Monitoring Single-vesicle Level Genetically-encoded Probes Spatio-temporal Resolution Total Internal Reflection Fluorescence Microscopy Evanescent Field High-contrast Images Vesicle Tracking Fusion Events SH-SY5Y Cells Synapto-pHluorin Transfection
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