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生物学
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Micromanipulation Techniques Allowing Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators
JoVE 杂志
生物学
This content is Free Access.
JoVE 杂志 生物学
Micromanipulation Techniques Allowing Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators

Micromanipulation Techniques Allowing Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators

DOI:
10.3791/57643-v

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12:52 min

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May 12, 2018

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1Division of Molecular Cell Biology, Zoological Institute,Technische Universität Braunschweig, 2Department of Cell Biology,Helmholtz Centre for Infection Research

Chapters

  • 00:05Title
  • 00:50Treatment of Cells, Transfection, and Seeding onto Coverslips
  • 02:54Assembly of Microscopy Imaging Chamber
  • 03:43Microinjection Procedure
  • 05:02FRAP Procedure
  • 05:32Photoactivation Procedure
  • 06:26Data Analysis
  • 09:42Results: Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators
  • 11:27Conclusion

Summary

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We describe how micro- and photomanipulation techniques such as FRAP and photoactivation enable the determination of motility parameters and the spatiotemporal dynamics of proteins within migrating cells. Experimental readouts include subcellular dynamics and turnover of motility regulators or of the underlying actin cytoskeleton.

Tags

Keywords: MicromanipulationCytoskeletal RegulatorsFRAPPhotoactivationB16-F1 CellsNIH 3T3 FibroblastsLamininFibronectinTransfectionMicroscopy

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