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Verfasst von:Cremer, Christoph [VerfasserIn]   i
 Schock, Florian [VerfasserIn]   i
 Birk, Udo J. [VerfasserIn]   i
Titel:Super-resolution microscopy approaches to nuclear nanostructure imaging
Verf.angabe:Christoph Cremer, Aleksander Szczurek, Florian Schock, Amine Gourram, Udo Birk
E-Jahr:2017
Jahr:6 April 2017
Umfang:22 S.
Fussnoten:Gesehen am 02.10.2018
Titel Quelle:Enthalten in: Methods
Ort Quelle:Orlando, Fla. : Academic Press, 1990
Jahr Quelle:2017
Band/Heft Quelle:123(2017), Seite 11-32
ISSN Quelle:1095-9130
Abstract:The human genome has been decoded, but we are still far from understanding the regulation of all gene activities. A largely unexplained role in these regulatory mechanisms is played by the spatial organization of the genome in the cell nucleus which has far-reaching functional consequences for gene regulation. Until recently, it appeared to be impossible to study this problem on the nanoscale by light microscopy. However, novel developments in optical imaging technology have radically surpassed the limited resolution of conventional far-field fluorescence microscopy (ca. 200nm). After a brief review of available super-resolution microscopy (SRM) methods, we focus on a specific SRM approach to study nuclear genome structure at the single cell/single molecule level, Spectral Precision Distance/Position Determination Microscopy (SPDM). SPDM, a variant of localization microscopy, makes use of conventional fluorescent proteins or single standard organic fluorophores in combination with standard (or only slightly modified) specimen preparation conditions; in its actual realization mode, the same laser frequency can be used for both photoswitching and fluorescence read out. Presently, the SPDM method allows us to image nuclear genome organization in individual cells down to few tens of nanometer (nm) of structural resolution, and to perform quantitative analyses of individual small chromatin domains; of the nanoscale distribution of histones, chromatin remodeling proteins, and transcription, splicing and repair related factors. As a biomedical research application, using dual-color SPDM, it became possible to monitor in mouse cardiomyocyte cells quantitatively the effects of ischemia conditions on the chromatin nanostructure (DNA). These novel “molecular optics” approaches open an avenue to study the nuclear landscape directly in individual cells down to the single molecule level and thus to test models of functional genome architecture at unprecedented resolution.
DOI:doi:10.1016/j.ymeth.2017.03.019
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

Volltext ; Verlag: http://dx.doi.org/10.1016/j.ymeth.2017.03.019
 Volltext: http://www.sciencedirect.com/science/article/pii/S1046202317300130
 DOI: https://doi.org/10.1016/j.ymeth.2017.03.019
Datenträger:Online-Ressource
Sprache:eng
Sach-SW:(Single molecule) localization microscopy (SMLM)
 Nanoscopy
 Nuclear structure
 PALM
 SPDM
 STORM/dSTORM
 Super-resolution light microscopy (SRM)
K10plus-PPN:1581506406
Verknüpfungen:→ Zeitschrift

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