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Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)
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Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

DOI:
10.3791/54774-v

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11:57 min

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December 01, 2016

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,
1Department of Biology,South University of Science and Technology of China, Shenzhen, 2Mechanobiology Institute, Singapore, 3Department of Biomedical Engineering,National University of Singapore

Chapters

  • 00:05Title
  • 00:55Imaging Specimen Preparation
  • 03:11Sample Placement and iPALM Alignment
  • 05:29Calibration of the iPALM Setup
  • 07:47Data Acquisition
  • 09:09Results: Actin Cytoskeleton of Adherent Mammalian Cells Visualized by iPALM with Sub-20 nm Spatial Resolution in 3D
  • 10:55Conclusion

Summary

Automatic Translation

Automatic Translation

We present a protocol for the application of interferometric PhotoActivated Localization Microscopy (iPALM), a 3-dimensional single-molecule localization super resolution microscopy method, to the imaging of the actin cytoskeleton in adherent mammalian cells. This approach allows light-based visualization of nanoscale structural features that would otherwise remain unresolved by conventional diffraction-limited optical microscopy.

Tags

Keywords: 3D Super Resolution MicroscopyIPALMF-actin FilamentsUltrastructural VisualizationSingle Molecule LocalizationCell Sample PreparationImaging BufferCover Glass AssemblyEpoxy SealingNewton’s Rings Pattern

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