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The intrinsic polarity of cells can be primarily attributed to two factors- i) the asymmetric accumulation of mobile components such are regulatory molecules and subcellular components across the cell and ii) the orientation of polar cytoskeletal filaments that make up the cytoskeletal networks, specifically microfilaments, and microtubules arranged along the axis of polarity. Interactions between the cytoskeletal filaments are crucial for the establishment and maintenance of the polar nature...
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Quantifying Cytoskeleton Dynamics Using Differential Dynamic Microscopy
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Visualizing and Quantifying In Vivo Cortical Cytoskeleton Structure and Dynamics.

Amparo Rosero1,2, Denisa Oulehlová1,3, Viktor Žárský1,3

  • 1Department of Experimental Plant Biology, Faculty of Science, Charles University, Prague, Czech Republic.

Methods in Molecular Biology (Clifton, N.J.)
|June 1, 2019
PubMed
Summary

Researchers visualize plant cell shaping using advanced microscopy and ImageJ software. This allows detailed study of the cortical cytoskeleton

Keywords:
ActinCLSMFluorescent proteinsImage analysisImageJMicrotubulesSDCMVAEM

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Area of Science:

  • Plant Cell Biology
  • Cytoskeletal Dynamics
  • Microscopy Techniques

Background:

  • The cortical microtubule and actin cytoskeleton are crucial for plant cell morphogenesis.
  • Studying these dynamic structures in living cells requires advanced imaging methods.

Purpose of the Study:

  • To detail noninvasive in vivo methods for observing the plant cortical cytoskeleton.
  • To highlight the utility of advanced microscopy and image analysis for cytoskeletal research.

Main Methods:

  • Utilizing transgenic plants with fluorescent protein markers for actin and microtubules.
  • Employing advanced microscopy: confocal laser scanning microscopy (CLSM), spinning disk confocal microscopy (SDCM), and variable angle epifluorescence microscopy (VAEM).
  • Applying ImageJ software for quantitative analysis of microscopic image sequences.

Main Results:

  • Achieved unprecedented spatial and temporal resolution in imaging the cortical cytoskeleton of living plant cells.
  • Enabled quantitative extraction of data on cytoskeletal architecture and dynamics.
  • Provided insights into the roles of microtubule and actin networks in cell shaping.

Conclusions:

  • Advanced microscopy and image analysis tools offer powerful capabilities for studying plant cytoskeletal dynamics.
  • Noninvasive in vivo imaging is essential for understanding the real-time function of the cytoskeleton in plant cells.