Spatial Statistics of Three-Dimensional Growth Dynamics of Spindle Microtubules

Methods Mol Biol. 2025:2872:51-72. doi: 10.1007/978-1-0716-4224-5_4.

Abstract

The latest high-resolution 3D live-cell imaging technology, lattice light-sheet microscopy (LLSM), has successfully tracked the dynamics of microtubule growth throughout the entire mitotic spindle with unparalleled precision. By using green fluorescent protein-labeled end-binding protein 1 (EB1-GFP) as a marker for growing microtubule ends, LLSM has generated an extensive collection of multidimensional datasets mapping the positions and trajectories of these growing microtubule ends. Processing this data requires statistical analysis in three-dimensional space. This chapter describes the spatial statistical methods developed for this purpose, illustrated with practical examples. Finally, we discuss future prospects for analyzing complex, large-scale image data.

Keywords: EB1; LLSM; Lattice light-sheet microscopy; Microtubule dynamics; Mitosis; Spatial statistics; Spindle.

MeSH terms

  • Green Fluorescent Proteins / metabolism
  • Humans
  • Image Processing, Computer-Assisted / methods
  • Imaging, Three-Dimensional* / methods
  • Microscopy, Fluorescence / methods
  • Microtubule-Associated Proteins / metabolism
  • Microtubules* / metabolism
  • Spindle Apparatus* / metabolism

Substances

  • Green Fluorescent Proteins
  • Microtubule-Associated Proteins