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FRET Imaging in Three-dimensional Hydrogels
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JoVE Journal Biology
FRET Imaging in Three-dimensional Hydrogels

FRET Imaging in Three-dimensional Hydrogels

DOI:
10.3791/54135-v

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09:47 min

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August 01, 2016

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, ,
1Department of Oral Biology, Center for Craniofacial Regeneration, McGowan Institute of Regenerative Medicine,University of Pittsburgh, 2Department of Bioengineering, Center for Craniofacial Regeneration, McGowan Institute of Regenerative Medicine,University of Pittsburgh, 3Laerdal AS

Chapters

  • 00:05Title
  • 00:47Hydrogel Preparation
  • 02:303D FRET Imaging
  • 04:35FRET Ratio Calculation
  • 07:11Results: FRET Imaging of Forskolin-treated Cells in Hydrogels
  • 09:08Conclusion

Summary

Automatic Translation

Automatic Translation

Förster resonance energy transfer (FRET) imaging is a powerful tool for real-time cell biology studies. Here a method for FRET imaging cells in physiologic three-dimensional (3D) hydrogel microenvironments using conventional epifluorescence microscopy is presented. An analysis for ratiometric FRET probes that yields linear ratios over the activation range is described.

Tags

Keywords: FRET Imaging3D HydrogelsIntracellular SignalingDevelopmental BiologyPharmacologyRegenerative MedicineBiomaterialsDrug ScreeningWide Field MicroscopyHydrogel CastingCell SuspensionCrosslinkingUV ExposurePDMS DishPhenol-free MediumMicroscope ImagingFRET Ratio

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