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Meiosis pp 265-276 | Cite as

Microscopy Methods for Analysis of Spindle Dynamics in Meiotic Drosophila Spermatocytes

  • Matthew S. SavoianEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1471)

Abstract

The spindle is a microtubule-based structure whose remodeling is required for partitioning the chromosomes and cytoplasm during meiosis. Characterizing microtubule behavior is fundamental to understanding how these tubulin polymers contribute to successful cell division. Here, a procedure is described for the imaging and analysis of spindle microtubule dynamics in cultures of living Drosophila melanogaster primary spermatocytes expressing tubulin tagged with enhanced green fluorescent protein. It employs time-lapse scanning confocal microscopy and the photobleaching of fiduciary marks onto fluorescently tagged microtubules. These labels are subsequently used to determine the sites and rates of kinetochore fiber growth and shrinkage during metaphase. This method can be readily applied to different microtubule populations, meiotic stages, and genetic backgrounds.

Key words

Meiosis Kinetochore fiber Microtubule flux Fluorescent protein Time-lapse imaging Live cell imaging Photobleaching 

Abbreviations

DIC

Differential interference contrast

EGFP

Enhanced green fluorescent protein

k-fiber

Kinetochore fiber

MT

Microtubule

NA

Numerical aperture

PMT

Photomultiplier tube

ROI

Region of interest

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Institute of Fundamental SciencesMassey UniversityPalmerston NorthNew Zealand

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