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Spindle assembly on immobilized chromatin micropatterns.
Pugieux, Céline; Dmitrieff, Serge; Tarnawska, Katarzyna; Nédélec, François.
Afiliação
  • Pugieux C; Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
  • Dmitrieff S; Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
  • Tarnawska K; Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
  • Nédélec F; Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany. Electronic address: nedelec@embl.de.
Methods Enzymol ; 540: 435-48, 2014.
Article em En | MEDLINE | ID: mdl-24630121
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cromatina / Proteínas Imobilizadas / Fuso Acromático Limite: Animals Idioma: En Revista: Methods Enzymol Ano de publicação: 2014 Tipo de documento: Article País de afiliação: Alemanha

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cromatina / Proteínas Imobilizadas / Fuso Acromático Limite: Animals Idioma: En Revista: Methods Enzymol Ano de publicação: 2014 Tipo de documento: Article País de afiliação: Alemanha