Quantitatively Analyzing the Morphological Growth Dynamics of Bipolar and Non-bipolar Spindles Using Xenopus Egg Extract

Methods Mol Biol. 2025:2872:117-130. doi: 10.1007/978-1-0716-4224-5_8.

Abstract

The bipolar shape of the microtubule-based spindle is a pivotal morphological phenotype for accurate chromosome segregation during cell division. However, existing descriptions of spindle morphogenesis remain largely qualitative. Here, we introduce a method that provides a quantitative description of the morphological growth dynamics of spindles using Xenopus egg cytoplasmic extract and a computational image analysis pipeline. Our method allows for determining the characteristic growth dynamics of bipolar and non-bipolar spindles, comparing their spatiotemporal differences and changes in response to physical and biochemical perturbations.

Keywords: Large-field confocal live imaging; Machine learning; Morphodynamics; Quantitative shape analysis; Spindle assembly; Xenopus egg extract.

MeSH terms

  • Animals
  • Cell Extracts
  • Image Processing, Computer-Assisted / methods
  • Microtubules / metabolism
  • Ovum / cytology
  • Ovum / growth & development
  • Spindle Apparatus* / metabolism
  • Xenopus / growth & development
  • Xenopus laevis / growth & development

Substances

  • Cell Extracts