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In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy
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Biologia
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JoVE Journal Biologia
In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

DOI:
10.3791/2247-v

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14:26 min

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January 19, 2011

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, ,
1Department of Physics,University of Wisconsin – Milwaukee, 2Department of Biological Sciences,University of Wisconsin – Milwaukee

Capitoli

  • 00:00Titolo
  • 01:46Introduction
  • 03:04Calibrating the Spectrally Resolved Two-photon Microscope
  • 08:58Collecting Data on Biological Samples of Interest
  • 12:36Protein Interactions Determined by Spectrally Resolved Two-photon Microscopy
  • 14:03Conclusion

Summary

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By employing a spectrally resolved two-photon microscopy imaging system, pixel-level maps of Förster Resonance Energy Transfer (FRET) efficiencies are obtained for cells expressing membrane receptors hypothesized to form homo-oligomeric complexes. From the FRET efficiency maps, we are able to estimate stoichiometric information about the oligomer complex under study.

Tags

In Vivo QuantificationG Protein Coupled Receptor InteractionsSpectrally Resolved Two-photon MicroscopyProtein InteractionsFRETFluorescent ProbesLiving CellsIndustrial ApplicationsBasic Biological KnowledgeDonor MoleculeAcceptor MoleculeLaser LightFluorescence EmissionSpectral PropertiesProtein Interactions DegreeCell ScanningMolecular CompositionAcquisition ProcessSpatial DeterminationQuantitative ValuesSpectrally Resolved Two-photon Microscope

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