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Updated: Jun 10, 2026
Micromanipulation Techniques Allowing Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators
Published on: May 12, 2018
The use of fluorescence redistribution after photobleaching for analysis of cellular microtubule dynamics
Claire E Walczak1, Rania S Rizk, Sidney L Shaw
1Medical Sciences, Indiana University, Bloomington, Indiana 47405, USA.
Abstract:
Microtubules (MTs) are highly dynamic polymers that serve as tracks for vesicular movement during interphase and as structural components of the mitotic spindle, which is used to segregate the genetic material. MT dynamics are highly regulated wherein MTs turnover differentially between interphase and mitosis. Within the mitotic spindle, there are distinct classes of MTs with different dynamic properties. To understand how cellular proteins regulate the dynamics of MTs, it is necessary to have methods to assess their turnover properties. In this chapter we present approaches to assess MT dynamics in cultured mammalian cells using fluorescence redistribution after photobleaching. We include a discussion of cell culture and imaging conditions that maintain cell viability. We also provide an extensive discussion of both data collection and analysis that are utilized to estimate the turnover dynamics of MTs.
Insights
This study details methods for assessing microtubule (MT) dynamics in mammalian cells. Fluorescence redistribution after photobleaching (FRAP) is presented to analyze MT turnover and regulation.
Area of Science:
- Cell Biology
- Biophysics
Background:
- Microtubules (MTs) are dynamic polymers crucial for intracellular transport and cell division.
- MT dynamics are tightly regulated, varying between interphase and mitosis.
- Understanding MT regulation requires methods to assess turnover properties.
Purpose of the Study:
- To present methods for assessing microtubule dynamics in cultured mammalian cells.
- To provide guidance on cell culture, imaging, data collection, and analysis for MT turnover studies.
Main Methods:
- Fluorescence Redistribution After Photobleaching (FRAP) is the primary technique discussed.
- FRAP allows for the assessment of microtubule turnover rates.
- Optimized cell culture and imaging conditions are detailed to ensure cell viability.
Main Results:
- The chapter provides a comprehensive guide to applying FRAP for MT dynamics.
- Detailed protocols for data collection and analysis are included.
- The methods enable quantitative estimation of MT turnover.
Conclusions:
- FRAP is a valuable technique for studying microtubule dynamics.
- The presented approaches facilitate the investigation of how cellular proteins regulate MT turnover.
- This work supports research into the dynamic behavior of microtubules in various cellular processes.
