Fast and parallel nanoscale three-dimensional tracking of heterogeneous mammalian chromatin dynamics

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dc.citation.issueNumber6en_US
dc.citation.journalTitleMolecular Biology of the Cellen_US
dc.citation.volumeNumber33en_US
dc.contributor.authorGustavsson, Anna-Karinen_US
dc.contributor.authorGhosh, Rajarshi P.en_US
dc.contributor.authorPetrov, Petar N.en_US
dc.contributor.authorLiphardt, Jan T.en_US
dc.contributor.authorMoerner, W. E.en_US
dc.contributor.orgSmalley–Curl Instituteen_US
dc.contributor.orgInstitute of Biosciences and Bioengineeringen_US
dc.date.accessioned2022-08-04T14:53:24Zen_US
dc.date.available2022-08-04T14:53:24Zen_US
dc.date.issued2022en_US
dc.description.abstractChromatin organization and dynamics are critical for gene regulation. In this work we present a methodology for fast and parallel three-dimensional (3D) tracking of multiple chromosomal loci of choice over many thousands of frames on various timescales. We achieved this by developing and combining fluorogenic and replenishable nanobody arrays, engineered point spread functions, and light sheet illumination. The result is gentle live-cell 3D tracking with excellent spatiotemporal resolution throughout the mammalian cell nucleus. Correction for both sample drift and nuclear translation facilitated accurate long-term tracking of the chromatin dynamics. We demonstrate tracking both of fast dynamics (50 Hz) and over timescales extending to several hours, and we find both large heterogeneity between cells and apparent anisotropy in the dynamics in the axial direction. We further quantify the effect of inhibiting actin polymerization on the dynamics and find an overall increase in both the apparent diffusion coefficient D* and anomalous diffusion exponent α and a transition to more-isotropic dynamics in 3D after such treatment. We think that in the future our methodology will allow researchers to obtain a better fundamental understanding of chromatin dynamics and how it is altered during disease progression and after perturbations of cellular function.en_US
dc.identifier.citationGustavsson, Anna-Karin, Ghosh, Rajarshi P., Petrov, Petar N., et al.. "Fast and parallel nanoscale three-dimensional tracking of heterogeneous mammalian chromatin dynamics." <i>Molecular Biology of the Cell,</i> 33, no. 6 (2022) The American Society for Cell Biology: https://doi.org/10.1091/mbc.E21-10-0514.en_US
dc.identifier.digitalmbc-e21-10-0514en_US
dc.identifier.doihttps://doi.org/10.1091/mbc.E21-10-0514en_US
dc.identifier.urihttps://hdl.handle.net/1911/112975en_US
dc.language.isoengen_US
dc.publisherThe American Society for Cell Biologyen_US
dc.rightsThis article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial-Share Alike 4.0 International Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/4.0).en_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0).en_US
dc.titleFast and parallel nanoscale three-dimensional tracking of heterogeneous mammalian chromatin dynamicsen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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