Observation of live chromatin dynamics in cells via 3D localization microscopy using Tetrapod point spread functions

Biomed Opt Express. 2017 Nov 21;8(12):5735-5748. doi: 10.1364/BOE.8.005735. eCollection 2017 Dec 1.

Abstract

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.

Keywords: (070.0070) Fourier optics and signal processing; (170.1530) Cell analysis; (170.2520) Fluorescence microscopy; (170.6900) Three-dimensional microscopy.