title: Spindle assembly on immobilized chromatin micropatterns.
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.
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Céline
authors.initials: C
authors.lastname: Pugieux
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Serge
authors.initials: S
authors.lastname: Dmitrieff
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Tarnawska
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany. Electronic address: nedelec@embl.de.
authors.forename: François
authors.initials: F
authors.lastname: Nédélec
mesh_terms: D000818:Animals; D019921:Biotinylation; D002417:Cattle; D002843:Chromatin / Q000378:metabolism* / Q000648:ultrastructure; D007091:Image Processing, Computer-Assisted; D055767:Immobilized Proteins / Q000378:metabolism* / Q000648:ultrastructure; D012710:Serum Albumin, Bovine / Q000737:chemistry; D008941:Spindle Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D014981:Xenopus
chemical_list: D002843:Chromatin; D055767:Immobilized Proteins; D012710:Serum Albumin, Bovine
keywords: Automated image analysis; Chromatin micropatterns; Meiotic spindle movements; Spindle array