Tracking of multiple chromosomal loci in 3D throughout live mammalian cell nuclei

Anna-Karin Gustavsson; Rajarshi P. Ghosh; Petar N. Petrov; Jan Liphardt; W. E. Moerner

doi:10.1117/12.2578200

5 March 2021 Tracking of multiple chromosomal loci in 3D throughout live mammalian cell nuclei

Anna-Karin Gustavsson, Rajarshi P. Ghosh, Petar N. Petrov, Jan Liphardt, W. E. Moerner

Proceedings Volume 11650, Single Molecule Spectroscopy and Superresolution Imaging XIV; 116500J (2021) https://doi.org/10.1117/12.2578200
Event: SPIE BiOS, 2021, Online Only

Abstract

The organization and dynamics of the genome regulate DNA replication, DNA repair, and gene expression. Here, we developed a methodology for studies of 3D organization and dynamics of the nucleome over time scales ranging from milliseconds to hours, and length scales ranging from tens of nanometers to micrometers. We achieve unprecedented 3D track lengths throughout the mammalian nucleus by combining a novel labeling scheme using nanobody ArrayG/N fusions of dSpCas9 targeted to specific DNA loci; light sheet illumination for gentle imaging of live cells with high contrast; and point spread function engineering for parallel detection of multiple loci in 3D.

Conference Presentation

Citation Download Citation

Anna-Karin Gustavsson, Rajarshi P. Ghosh, Petar N. Petrov, Jan Liphardt, and W. E. Moerner "Tracking of multiple chromosomal loci in 3D throughout live mammalian cell nuclei", Proc. SPIE 11650, Single Molecule Spectroscopy and Superresolution Imaging XIV, 116500J (5 March 2021); https://doi.org/10.1117/12.2578200

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