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Time‐resolved Microscopy of Chromatin In Vitro and In Vivo ¶
Author(s) -
Davis Sara K.,
Bardeen Christopher J.
Publication year - 2005
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.2005.tb00224.x
Subject(s) - chromatin , in vivo , biophysics , histone , photobleaching , dna , fluorescence microscope , fluorescence recovery after photobleaching , in vitro , chemistry , fluorescence , biology , microbiology and biotechnology , physics , biochemistry , genetics , optics
In eukaryotic cell nuclei, double‐stranded DNA is found in the form of chromatin, a large fiber made up of DNA complexed to histone proteins. In this article, recent studies using fluorescence techniques to look at the dynamics of chromatin, both in vivo and in vitro , are reviewed. Two‐photon counter‐propagating fluorescence recovery after patterned photobleaching is used to examine chromatin fluctuations on lengthscales ranging from less than 100 nm to microns. By combining in vivo studies with data on isolated nuclei and by measuring how these fluctuations depend on variables like ionic strength and photochemical cross‐linking, it is demonstrated that the relatively large‐scale motions of chromatin observed in vivo are consistent with smaller scale modifications of the histone‐DNA interaction. This connection may provide a means to use conformational dynamics as an in vivo probe of the biochemical events involved in gene expression.