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Quantification of Intracellular Mitochondrial Displacements in Response to Nanomechanical Forces

  • Yaron R. Silberberg
  • Andrew E. Pelling
Part of the Methods in Molecular Biology book series (MIMB, volume 991)

Abstract

Mechanical stress affects various aspects of cell behavior, including cell growth, morphology, differentiation, and genetic expression. Here, we describe a method to quantify the intracellular mechanical response to an extracellular mechanical perturbation, specifically the displacement of mitochondria. A combined fluorescent-atomic force microscope (AFM) was used to simultaneously produce well-defined nanomechanical stimulation to a living cell while optically recording the real-time displacement of fluorescently labeled mitochondria. A single-particle tracking (SPT) approach was then applied in order to quantify the two-dimensional displacement of mitochondria in response to local forces.

Key words

Atomic force microscopy Mitochondria Single-particle tracking Nanomechanics Mechanotransduction Force transmission 

Notes

Acknowledgments

Y.R.S. would like to thank the Japanese Society for the Promotion of Science (JSPS) for a post-doctoral fellowship grant. A.E.P. acknowledges generous support from the Canada Research Chairs program, the Province of Ontario Early Researcher Award, and the Natural Sciences and Engineering Research Council. The authors would like to gratefully acknowledge the tremendous support and mentorship of Professor Michael Horton (1948–2010) and his inspiration for this work.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Yaron R. Silberberg
    • 1
  • Andrew E. Pelling
    • 2
  1. 1.Biomedical Research Institute (BMRI), National Institute of Advanced Industrial Science and Technology (AIST)Kyoto UniversityKyotoJapan
  2. 2.Department of Physics, Department of Biology, Institute for Science, Society and PolicyUniversity of OttawaOttawaCanada

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