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細胞イメージングおよびデータ分析：SH-SY5Y神経芽腫細胞における神経伝達物質小胞のダイナミクスを評価するためにTIRFMとpH感受性GFP-プローブ
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Neurosciences
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Journal JoVE Neurosciences
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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細胞イメージングおよびデータ分析：SH-SY5Y神経芽腫細胞における神経伝達物質小胞のダイナミクスを評価するためにTIRFMとpH感受性GFP-プローブ

10,586 Views

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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

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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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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