Analysis of microtubule dynamics heterogeneity in cell culture

Anara Serikbaeva, Anna Tvorogova, Sholpan Kauanova, Ivan Vorobyev

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Microtubules (MTs) are dynamic components of the cytoskeleton playing an important role in a large number of cell functions. Individual MTs in living cells undergo stochastic switching between alternate states of growth, shortening and attenuated phase, a phenomenon known as tempered dynamic instability. Dynamic instability of MTs is usually analyzed by labeling MTs with +TIPs, namely, EB proteins. Tracking of +TIP trajectories allows analyzing MT growth in cells with a different density of MTs. Numerous labs now use +TIP to track growing MTs in a variety of cell cultures. However, heterogeneity of MT dynamics is usually underestimated, and rather small sampling for the description of dynamic instability parameters is often used. The strategy described in this chapter is the method for repetitive quantitative analysis of MT growth rate within the same cell that allows minimization of the variation in MT dynamics measurement. We show that variability in MT dynamics within a cell when using repeated measurements is significantly less than between different cells in the same chamber. This approach allows better estimation of the heterogeneity of cells’ responses to different treatments. To compare the effects of different MT inhibitors, the protocol using normalized values for MT dynamics and repetitive measurements for each cell is employed. This chapter provides detailed methods for analysis of MT dynamics in tissue cultures. We describe protocols for imaging MT dynamics by fluorescent microscopy, contrast enhancement technique, and MT dynamics analysis using triple color-coded display based on sequential subtraction analysis.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages181-204
Number of pages24
DOIs
Publication statusPublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1745
ISSN (Print)1064-3745

Keywords

  • Dual color-coded display
  • End-binding protein
  • Fluorescent microscopy
  • Microtubule dynamics

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Analysis of microtubule dynamics heterogeneity in cell culture'. Together they form a unique fingerprint.

  • Cite this

    Serikbaeva, A., Tvorogova, A., Kauanova, S., & Vorobyev, I. (2018). Analysis of microtubule dynamics heterogeneity in cell culture. In Methods in Molecular Biology (pp. 181-204). (Methods in Molecular Biology; Vol. 1745). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7680-5_11