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Single-Molecule Imaging of Nuclear Transport
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A user-interactive algorithm quantifying nuclear pore complex distribution within the nuclear lamina network in

John S Y Lim1, Graham D Wright1, Brian Burke2

  • 1Institute of Medical Biology, Agency for Science, Technology and Research, 8A Biomedical Grove, #06-06 Immunos, Singapore 138648, Singapore; Skin Research Institute of Singapore, Agency for Science, Technology and Research, Singapore.

Methods (San Diego, Calif.)
|October 1, 2018
PubMed
Summary
This summary is machine-generated.

Super-resolution microscopy enables precise analysis of the nuclear envelope (NE). A new algorithm quantifies nuclear pore complex (NPC) localization within nuclear lamin networks in mammalian cells.

Keywords:
FijiNuclear envelopeNuclear laminNuclear pore complexSuper-resolution microscopy

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Area of Science:

  • Cell Biology
  • Microscopy
  • Biophysics

Background:

  • Mammalian nuclear envelope (NE) components like nuclear lamina and nuclear pore complexes (NPCs) are difficult to analyze quantitatively with conventional fluorescence microscopy due to sub-diffraction limit sizes.
  • Super-resolution microscopy (SRM) has overcome these limitations, enabling visualization of NE structures at the nanoscale in intact cells.

Purpose of the Study:

  • To develop and validate a computational tool for quantitative analysis of NE components.
  • To enable precise localization of nuclear pore complexes (NPCs) relative to nuclear lamin distribution.

Main Methods:

  • Utilized single molecule localization microscopy (SMLM) for high-resolution imaging of the nuclear envelope.
  • Developed a novel algorithm to identify NPC centroids and quantify their spatial relationship with lamin filaments.
  • Applied the algorithm for semi-automatic, nanoscale comparison of cellular component distribution.

Main Results:

  • The algorithm accurately identifies NPC centroids and their localization within the nuclear lamin network.
  • Quantitative comparison of NPC distribution across cells with varying lamin complements is now feasible.
  • The computational tool provides fast and accurate nanometre-scale quantification.

Conclusions:

  • Super-resolution microscopy combined with advanced algorithms offers powerful quantitative insights into nuclear envelope organization.
  • This approach facilitates comparative studies of NE component distribution and dynamics.
  • The developed algorithm serves as a versatile tool for nanoscale biological imaging analysis.