Quantification of Intracellular Mitochondrial Displacements in Response to Nanomechanical Forces
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 wordsAtomic force microscopy Mitochondria Single-particle tracking Nanomechanics Mechanotransduction Force transmission
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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