Spindle assembly on immobilized chromatin micropatterns

Methods Enzymol. 2014;540:435-48. doi: 10.1016/B978-0-12-397924-7.00024-8.

Abstract

We describe a method to assemble meiotic spindles on immobilized micropatterns of chromatin built on a first layer of biotinylated BSA deposited by microcontact printing. Such chromatin patterns routinely produce bipolar spindles with a yield of 60%, and offer the possibility to follow spindle assembly dynamics, from the onset of nucleation to the establishment of a quasi steady state. Hundreds of spindles can be recorded in parallel for different experimental conditions. We also describe the semi-automated image analysis pipeline, which is used to analyze the assembly kinetics of spindle arrays, or the final morphological diversity of the spindles.

Keywords: Automated image analysis; Chromatin micropatterns; Meiotic spindle movements; Spindle array.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotinylation
  • Cattle
  • Chromatin / metabolism*
  • Chromatin / ultrastructure
  • Image Processing, Computer-Assisted
  • Immobilized Proteins / metabolism*
  • Immobilized Proteins / ultrastructure
  • Serum Albumin, Bovine / chemistry
  • Spindle Apparatus / metabolism*
  • Spindle Apparatus / ultrastructure
  • Xenopus

Substances

  • Chromatin
  • Immobilized Proteins
  • Serum Albumin, Bovine