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Related Experiment Videos

Three-Dimensional Super-Resolution in Eukaryotic Cells Using the Double-Helix Point Spread Function.

Alexander R Carr1, Aleks Ponjavic1, Srinjan Basu2

  • 1Department of Chemistry, University of Cambridge, Cambridge, United Kingdom.

Biophysical Journal
|April 14, 2017
PubMed
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This summary is machine-generated.

This study extends double-helix point spread function (DHPSF) imaging to visualize whole eukaryotic cells in 3D. This breakthrough enables detailed tracking of protein organization and dynamics in complex cellular environments.

Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Single-molecule localization microscopy (SMLM) visualizes nanoscale protein organization and dynamics.
  • Existing SMLM methods are limited in 3D imaging of thick biological samples.
  • The double-helix point spread function (DHPSF) offers 3D localization but was previously restricted to small cells.

Purpose of the Study:

  • To adapt and demonstrate DHPSF imaging for 3D visualization of entire eukaryotic cells.
  • To overcome limitations of DHPSF in imaging thicker biological specimens.
  • To apply DHPSF for studying large-scale cellular dynamics and protein trafficking.

Main Methods:

  • Optimized DHPSF imaging by matching refractive indices of immersion liquid and sample media.

Related Experiment Videos

  • Imaged whole eukaryotic cell volumes up to 15 micrometers thick in 3-5 imaging planes.
  • Utilized single-particle tracking to analyze protein dynamics.

    • Main Results:

    • Successfully demonstrated DHPSF imaging in thick eukaryotic cells.
    • Visualized large-scale membrane reorganization in human T cells post-receptor triggering.
    • Tracked dynamics of membrane, cytoplasmic, and nuclear mammalian proteins in various cell types.

    Conclusions:

    • DHPSF imaging is now applicable to 3D visualization of whole eukaryotic cells.
    • This technique provides new insights into cellular processes and protein dynamics in complex environments.
    • DHPSF imaging advances the study of cellular organization and function in mammalian cells.