JoVE Journal

Biology

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

10.0K Views

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12:52 min

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May 12th, 2018

DOI :

10.3791/57643-v

May 12th, 2018

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Georgi Dimchev¹^,², Klemens Rottner¹^,²

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

Summary

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.

Explore More Videos

Micromanipulation

Cytoskeletal Regulators

Photoactivation

NIH 3T3 Fibroblasts

Chapters in this video

0:05

Title

0:50

Treatment of Cells, Transfection, and Seeding onto Coverslips

2:54

Assembly of Microscopy Imaging Chamber

3:43

Microinjection Procedure

5:02

FRAP Procedure

5:32

Photoactivation Procedure

6:26

Data Analysis

9:42

Results: Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators

11:27

Conclusion

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