Fast and parallel nanoscale three-dimensional tracking of heterogeneous mammalian chromatin dynamics.

ABSTRACT: Chromatin organization and dynamics are critical for gene regulation. In this work we present a methodology for fast and parallel three-dimensional (3D) tracking of multiple chromosomal loci of choice over many thousands of frames on various timescales. We achieved this by developing and combining fluorogenic and replenishable nanobody arrays, engineered point spread functions, and light sheet illumination. The result is gentle live-cell 3D tracking with excellent spatiotemporal resolution throughout the mammalian cell nucleus. Correction for both sample drift and nuclear translation facilitated accurate long-term tracking of the chromatin dynamics. We demonstrate tracking both of fast dynamics (50 Hz) and over timescales extending to several hours, and we find both large heterogeneity bet

SUBMITTER: Gustavsson AK 

PROVIDER: S-EPMC9265149 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature