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Real-time Imaging of Plant Cell Surface Dynamics with Variable-angle Epifluorescence Microscopy
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Real-time Imaging of Plant Cell Surface Dynamics with Variable-angle Epifluorescence Microscopy

Real-time Imaging of Plant Cell Surface Dynamics with Variable-angle Epifluorescence Microscopy

DOI:
10.3791/53437-v

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06:31 min

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December 12, 2015

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1Graduate School of Frontier Sciences,The University of Tokyo

Chapters

  • 00:05Title
  • 00:35Cotyledon specimen Sky Drop Mounting
  • 02:35Variable-angle Epifluorescence Microscopy (VAEM) Observation and Movie Acquisition
  • 03:42GFP-labeled Dot Residence Time Quantification by Kymograph Analysis
  • 05:19Results: Representative GFP-PATROL1 Dot Residence Time Analysis by Kymography
  • 06:02Conclusion

Summary

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Automatic Translation

The goal of this protocol is to demonstrate how to monitor fluorescently-tagged protein dynamics on plant cell surfaces with variable-angle epifluorescence microscopy, showing blinking dots of GFP-tagged PATROL1, a membrane trafficking protein, in the cell cortex of the stomatal complex in Arabidopsis thaliana.

Tags

Keywords: Cell Surface DynamicsVariable-angle Epifluorescence MicroscopyVAEMReal-time ImagingPlant Cell BiologyMembrane TraffickingCytoskeletal RearrangementFluorescently-labeled ProteinsPATROL1Stomatal MovementPlasma MembraneArabidopsis ThalianaKymograph AnalysisTime-lapse Imaging

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