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Fluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Fluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

Fluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

DOI:
10.3791/2568-v

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06:35 min

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April 16, 2011

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, , ,
1National Institute on Deafness and Other Communication Disorders,National Institutes of Health, Bethesda

Chapters

  • 00:05Title
  • 00:43Introduction
  • 01:15Neuron Transfection
  • 02:44FRAP Experiment on a Spine
  • 04:40Data Analysis
  • 05:28Representative Results: FRAP Measurements of EGFP Fluorescence in a Spine from a Cultured Hippocampal Neuron
  • 06:13Conclusion

Summary

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FRAP has been used to quantify the mobility of Green Fluorescence Protein (GFP)-tagged proteins in cultured cells. We examined the mobile/immobile fractions of the GFP by analyzing the fluorescence recovery percentage after photobleaching. In this study, FRAP was performed at spines of hippocampal neurons.

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Fluorescence Recovery After Photobleaching (FRAP)GFP-tagged ProteinsDendritic SpinesCultured Hippocampal NeuronsMobilityExcitation LaserFluorescence Recovery RateProtein Turnover RateEGFP VectorZeiss 710 Confocal MicroscopePhotobleachingTime Lapse ImagesMobile FractionImmobile FractionImageJ SoftwareGraphpad Software

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