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

Spindle assembly on immobilized chromatin micropatterns.

Author information

1
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
2
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany. Electronic address: nedelec@embl.de.

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

[Indexed for MEDLINE]

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