Localization microscopy of DNA in situ using Vybrant(®) DyeCycle™ Violet fluorescent probe: A new approach to study nuclear nanostructure at single molecule resolution

Experimental Cell Research
Dominika J Zurek-BiesiadaChristoph Cremer

Abstract

Higher order chromatin structure is not only required to compact and spatially arrange long chromatids within a nucleus, but have also important functional roles, including control of gene expression and DNA processing. However, studies of chromatin nanostructures cannot be performed using conventional widefield and confocal microscopy because of the limited optical resolution. Various methods of superresolution microscopy have been described to overcome this difficulty, like structured illumination and single molecule localization microscopy. We report here that the standard DNA dye Vybrant(®) DyeCycle™ Violet can be used to provide single molecule localization microscopy (SMLM) images of DNA in nuclei of fixed mammalian cells. This SMLM method enabled optical isolation and localization of large numbers of DNA-bound molecules, usually in excess of 10(6) signals in one cell nucleus. The technique yielded high-quality images of nuclear DNA density, revealing subdiffraction chromatin structures of the size in the order of 100nm; the interchromatin compartment was visualized at unprecedented optical resolution. The approach offers several advantages over previously described high resolution DNA imaging methods, including high spec...Continue Reading

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Citations

Nov 13, 2015·Proceedings of the National Academy of Sciences of the United States of America·Kirti PrakashUdo J Birk
Jul 26, 2016·Frontiers in Genetics·Aleksander SzczurekChristoph Cremer
May 28, 2019·The FEBS Journal·Jianquan Xu, Yang Liu
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Aug 26, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Paul J SmithRachel J Errington
Jan 27, 2019·Chemical Communications : Chem Comm·Xiaodong ZhangYi Xiao
Nov 7, 2015·Genome Biology·Ina KirmesGeorge Reid
Apr 27, 2021·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Kirti Prakash

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