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Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

DOI:
10.3791/54466-v

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07:26 min

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October 15, 2016

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1Department of Neuroscience,University of Wisconsin-Madison

Chapters

  • 00:05Title
  • 01:03Membrane Sheet Preparation and Fixation
  • 02:53Photoactivation Localization Microscopy (PALM) Image Acquisition
  • 05:40Results: Images of PI(4,5)P2 and iRFP-PAmCherry1-PHPLCδ1 Labeled INS-1 Cells
  • 06:56Conclusion

Summary

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Automatic Translation

PI(4,5)P2 regulates various cellular functions, but its nanoscale organization in the cell plasma membrane is poorly understood. By labeling PI(4,5)P2 with a dual-color fluorescent probe fused to the Pleckstrin Homology domain, we describe a novel approach to study the PI(4,5)P2 spatial distribution in the plasma membrane at the nanometer scale.

Tags

Keywords: Single-molecule Super-resolution ImagingPhosphatidylinositol 45-bisphosphatePlasma MembraneFluorescent ProbesNanoscale DistributionPhosphoinositide SignalingHigh Spatial ResolutionDetergent-freePlasma Membrane Sheet PreparationPhosphoinositide LabelingSingle-molecule Fluorescence ImagingSingle-molecule LocalizationTIRF MicroscopySMLM

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