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

Details

Serval ID
serval:BIB_C78C62B525B3
Type
Article: article from journal or magazin.
Collection
Publications
Title
Localization microscopy of DNA in situ using Vybrant(®) DyeCycle™ Violet fluorescent probe: A new approach to study nuclear nanostructure at single molecule resolution.
Journal
Experimental cell research
Author(s)
Żurek-Biesiada D., Szczurek A.T., Prakash K., Mohana G.K., Lee H.K., Roignant J.Y., Birk U.J., Dobrucki J.W., Cremer C.
ISSN
1090-2422 (Electronic)
ISSN-L
0014-4827
Publication state
Published
Issued date
01/05/2016
Peer-reviewed
Oui
Volume
343
Number
2
Pages
97-106
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
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 specificity, an ability to record images using a single wavelength excitation, and a higher density of single molecule signals than reported in previous SMLM studies. The method is compatible with DNA/multicolor SMLM imaging which employs simple staining methods suited also for conventional optical microscopy.
Keywords
Animals, Cell Nucleus/metabolism, Cercopithecus aethiops, Chromosomes/metabolism, DNA/metabolism, Drosophila melanogaster, Fluorescent Dyes/metabolism, Microscopy, Fluorescence/methods, Nanostructures/chemistry, Single Molecule Imaging/methods, Vero Cells, Chromatin structure, DNA dyes, Photoconversion, SMLM, Vybrant DyeCycle Violet, dSTORM
Pubmed
Web of science
Create date
28/10/2019 12:56
Last modification date
29/10/2019 6:26
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