Replication Labeling Methods for Super-Resolution Imaging of Chromosome Territories and Chromatin Domains
Publikation: Bidrag til bog/antologi/rapport › Bidrag til bog/antologi › Forskning › fagfællebedømt
Continuing progress in super-resolution microscopy enables the study of sub-chromosomal chromatin organization in single cells with unprecedented detail. Here we describe refined methods for pulse-chase replication labeling of individual chromosome territories (CTs) and replication domain units in mammalian cell nuclei, with specific focus on their application to three-dimensional structured illumination microscopy (3D-SIM). We provide detailed protocols for highly efficient electroporation-based delivery or scratch loading of cell-impermeable fluorescent nucleotides for live-cell studies. Furthermore, we describe the application of (2'S)-2'-deoxy-2'-fluoro-5-ethynyluridine (F-ara-EdU) and 5-vinyl-2'-deoxyuridine (VdU) for the in situ detection of segregated chromosome territories and sister chromatids with minimized cytotoxic side effects.
Originalsprog | Engelsk |
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Titel | Chromosome Architecture |
Redaktører | Mark C. Leake |
Antal sider | 18 |
Forlag | Springer |
Publikationsdato | 2022 |
Udgave | 2 |
Sider | 111-128 |
ISBN (Trykt) | 978-1-0716-2223-0, 978-1-0716-2220-9 |
ISBN (Elektronisk) | 978-1-0716-2221-6 |
DOI | |
Status | Udgivet - 2022 |
Navn | Methods in Molecular Biology |
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Vol/bind | 2476 |
ISSN | 1064-3745 |
Bibliografisk note
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
ID: 367708001