High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain
Name
document.pdf
Description
Published version
Size
771.84 KB
Format
Adobe PDF
Checksum (MD5)
d1cc60bf194baa26f780cf67996efe83
Author(s)
Makhija, Ekta
Jagielska, Anna
Van Vliet, Krystyn J
Date Issued
2019
Journal
Journal of Visualized Experiments
Publisher
MyJove Corporation
Version
Final published version
Abstract
© 2019 Journal of Visualized Experiments. Extracellular mechanical strain is known to elicit cell phenotypic responses and has physiological relevance in several tissue systems. To capture the effect of applied extracellular tensile strain on cell populations in vitro via biochemical assays, a device has previously been designed which can be fabricated simply and is small enough to fit inside tissue culture incubators, as well as on top of microscope stages. However, the previous design of the polydimethylsiloxane substratum did not allow high-resolution subcellular imaging via oil-immersion objectives. This work describes a redesigned geometry of the polydimethylsiloxane substratum and a customized imaging setup that together can facilitate high-resolution subcellular imaging of live cells while under applied strain. This substratum can be used with the same, earlier designed device and, hence, has the same advantages as listed above, in addition to allowing high-resolution optical imaging. The design of the polydimethylsiloxane substratum can be improved by incorporating a grid which will facilitate tracking the same cell before and after the application of strain. Representative results demonstrate high-resolution time-lapse imaging of fluorescently labeled nuclei within strained cells captured using the method described here. These nuclear dynamics data give insights into the mechanism by which applied tensile strain promotes differentiation of oligodendrocyte progenitor cells.
MIT Department
Massachusetts Institute of Technology. Department of Materials Science and Engineering
Massachusetts Institute of Technology. Department of Biological Engineering
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DOI of Published Version
https://doi.org/10.3791/59474
