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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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10.7K Views

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13:47 min

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January 29th, 2015

DOI :

10.3791/52267-v

January 29th, 2015

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Federica Daniele¹, Eliana S. Di Cairano¹, Stefania Moretti¹, Giovanni Piccoli², Carla Perego¹^,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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TIRFM

PH sensitive GFP probes

Neurotransmitter Vesicle Dynamics

SH SY5Y Neuroblastoma Cells

Total Internal Reflection Fluorescence Microscopy

Synapto pHluorin

Membrane Depolarization

Neurotransmitter Release

Fusion Events

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0:05

Title

3:32

Cell Imaging by Total Internal Reflection Fluorescence Microscopy (TIRFM)

6:08

Fluorescence Intensity Quantification

7:09

Photobleaching Correction and Threshold Determination

8:16

Selection of Fusion Events Using a Semiautomatic Procedure

10:01

Data Analysis

11:31

Results: Analysis of Synaptic Vesicles Fusion Under Resting and Stimulated (Potassium-induced Depolarization) Conditions Are Shown

12:37

Conclusion

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