Spindle assembly on immobilized chromatin micropatterns
2014
Authors: Pugieux C, Dmitrieff S, Tarnawska K, Nédélec F.
CellNetworks People: Nédélec François
Journal: Methods Enzymol. 2014;540:435-48

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