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TIRFM og pH-følsomme GFP-prober til Evaluer Neurotransmitter Vesikel Dynamics i SH-SY5Y neuroblastomaceller: Cell Imaging og Data Analysis
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JoVE Journal Neuroscience
TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis

TIRFM og pH-følsomme GFP-prober til Evaluer Neurotransmitter Vesikel Dynamics i SH-SY5Y neuroblastomaceller: Cell Imaging og Data Analysis

DOI:
10.3791/52267-v

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13:47 min

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January 29, 2015

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, , , ,
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

Chapters

  • 00:05Title
  • 03:32Cell Imaging by Total Internal Reflection Fluorescence Microscopy (TIRFM)
  • 06:08Fluorescence Intensity Quantification
  • 07:09Photobleaching Correction and Threshold Determination
  • 08:16Selection of Fusion Events Using a Semiautomatic Procedure
  • 10:01Data Analysis
  • 11:31Results: Analysis of Synaptic Vesicles Fusion Under Resting and Stimulated (Potassium-induced Depolarization) Conditions Are Shown
  • 12:37Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

Keywords: 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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