Open AccessObservation of live chromatin dynamics in cells via 3D localization microscopy using Tetrapod point spread functions
Author(s) -
Yoav Shechtman,
Anna-Karin Gustavsson,
Petar N. Petrov,
Elisa Dultz,
Maurice Y. Lee,
Karsten Weis,
W. E. Moerner
Publication year - 2017
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.8.005735
Subject(s) - euchromatin , chromatin , heterochromatin , budding yeast , saccharomyces cerevisiae , locus (genetics) , biology , microscopy , genetics , evolutionary biology , dna , yeast , optics , physics , gene
We report the observation of chromatin dynamics in living budding yeast ( Saccharomyces cerevisiae ) cells, in three-dimensions (3D). Using dual color localization microscopy and employing a Tetrapod point spread function, we analyze the spatio-temporal dynamics of two fluorescently labeled DNA loci surrounding the GAL locus. From the measured trajectories, we obtain different dynamical characteristics in terms of inter-loci distance and temporal variance; when the GAL locus is activated, the 3D inter-loci distance and temporal variance increase compared to the inactive state. These changes are visible in spite of the large thermally- and biologically-driven heterogeneity in the relative motion of the two loci. Our observations are consistent with current euchromatin vs. heterochromatin models.