In vivo and in situ replication labeling methods for super-resolution structured illumination microscopy of chromosome territories and chromatin domains
Recent advances in super-resolution microscopy enable the study of subchromosomal 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 mammali...
| Main Authors: | , , , |
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| Format: | Journal article |
| Language: | English |
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Springer New York
2016
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| _version_ | 1797092951578378240 |
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| author | Miron, E Innocent, C Heyde, S Schermelleh, L |
| author_facet | Miron, E Innocent, C Heyde, S Schermelleh, L |
| author_sort | Miron, E |
| collection | OXFORD |
| description | Recent advances in super-resolution microscopy enable the study of subchromosomal 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) for the in situ detection of segregated chromosome territories with minimized cytotoxic side effects. |
| first_indexed | 2024-03-07T03:53:21Z |
| format | Journal article |
| id | oxford-uuid:c206c601-7710-488f-9a8e-4dd6eeb4e9b0 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:53:21Z |
| publishDate | 2016 |
| publisher | Springer New York |
| record_format | dspace |
| spelling | oxford-uuid:c206c601-7710-488f-9a8e-4dd6eeb4e9b02022-03-27T06:05:59ZIn vivo and in situ replication labeling methods for super-resolution structured illumination microscopy of chromosome territories and chromatin domainsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:c206c601-7710-488f-9a8e-4dd6eeb4e9b0EnglishSymplectic Elements at OxfordSpringer New York2016Miron, EInnocent, CHeyde, SSchermelleh, LRecent advances in super-resolution microscopy enable the study of subchromosomal 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) for the in situ detection of segregated chromosome territories with minimized cytotoxic side effects. |
| spellingShingle | Miron, E Innocent, C Heyde, S Schermelleh, L In vivo and in situ replication labeling methods for super-resolution structured illumination microscopy of chromosome territories and chromatin domains |
| title | In vivo and in situ replication labeling methods for super-resolution structured illumination microscopy of chromosome territories and chromatin domains |
| title_full | In vivo and in situ replication labeling methods for super-resolution structured illumination microscopy of chromosome territories and chromatin domains |
| title_fullStr | In vivo and in situ replication labeling methods for super-resolution structured illumination microscopy of chromosome territories and chromatin domains |
| title_full_unstemmed | In vivo and in situ replication labeling methods for super-resolution structured illumination microscopy of chromosome territories and chromatin domains |
| title_short | In vivo and in situ replication labeling methods for super-resolution structured illumination microscopy of chromosome territories and chromatin domains |
| title_sort | in vivo and in situ replication labeling methods for super resolution structured illumination microscopy of chromosome territories and chromatin domains |
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