High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain
© 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 devic...
| Main Authors: | , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
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MyJove Corporation
2021
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| Online Access: | https://hdl.handle.net/1721.1/134816 |
| _version_ | 1811069378064547840 |
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| author | Makhija, Ekta Jagielska, Anna Van Vliet, Krystyn J |
| author2 | Massachusetts Institute of Technology. Department of Materials Science and Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Materials Science and Engineering Makhija, Ekta Jagielska, Anna Van Vliet, Krystyn J |
| author_sort | Makhija, Ekta |
| collection | MIT |
| description | © 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. |
| first_indexed | 2024-09-23T08:10:04Z |
| format | Article |
| id | mit-1721.1/134816 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T08:10:04Z |
| publishDate | 2021 |
| publisher | MyJove Corporation |
| record_format | dspace |
| spelling | mit-1721.1/1348162023-02-17T19:59:55Z High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain Makhija, Ekta Jagielska, Anna Van Vliet, Krystyn J Massachusetts Institute of Technology. Department of Materials Science and Engineering Massachusetts Institute of Technology. Department of Biological Engineering © 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. 2021-10-27T20:09:19Z 2021-10-27T20:09:19Z 2019 2019-09-24T18:02:51Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/134816 en 10.3791/59474 Journal of Visualized Experiments Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf MyJove Corporation Journal of Visualized Experiments (JOVE) |
| spellingShingle | Makhija, Ekta Jagielska, Anna Van Vliet, Krystyn J High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain |
| title | High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain |
| title_full | High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain |
| title_fullStr | High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain |
| title_full_unstemmed | High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain |
| title_short | High-resolution Imaging of Nuclear Dynamics in Live Cells under Uniaxial Tensile Strain |
| title_sort | high resolution imaging of nuclear dynamics in live cells under uniaxial tensile strain |
| url | https://hdl.handle.net/1721.1/134816 |
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