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The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes
Chromatin undergoes dramatic condensation and decondensation as cells transition between the different phases of the cell cycle. The organization of chromatin in chromosomes is still one of the key challenges in structural biology. Fluorescence lifetime imaging (FLIM), a technique which utilizes a f...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4985626/ https://www.ncbi.nlm.nih.gov/pubmed/27526631 http://dx.doi.org/10.1038/srep31417 |
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author | Estandarte, Ana Katrina Botchway, Stanley Lynch, Christophe Yusuf, Mohammed Robinson, Ian |
author_facet | Estandarte, Ana Katrina Botchway, Stanley Lynch, Christophe Yusuf, Mohammed Robinson, Ian |
author_sort | Estandarte, Ana Katrina |
collection | PubMed |
description | Chromatin undergoes dramatic condensation and decondensation as cells transition between the different phases of the cell cycle. The organization of chromatin in chromosomes is still one of the key challenges in structural biology. Fluorescence lifetime imaging (FLIM), a technique which utilizes a fluorophore’s fluorescence lifetime to probe changes in its environment, was used to investigate variations in chromatin compaction in fixed human chromosomes. Fixed human metaphase and interphase chromosomes were labeled with the DNA minor groove binder, DAPI, followed by measurement and imaging of the fluorescence lifetime using multiphoton excitation. DAPI lifetime variations in metaphase chromosome spreads allowed mapping of the differentially compacted regions of chromatin along the length of the chromosomes. The heteromorphic regions of chromosomes 1, 9, 15, 16, and Y, which consist of highly condensed constitutive heterochromatin, showed statistically significant shorter DAPI lifetime values than the rest of the chromosomes. Differences in the DAPI lifetimes for the heteromorphic regions suggest differences in the structures of these regions. DAPI lifetime variations across interphase nuclei showed variation in chromatin compaction in interphase and the formation of chromosome territories. The successful probing of differences in chromatin compaction suggests that FLIM has enormous potential for application in structural and diagnostic studies. |
format | Online Article Text |
id | pubmed-4985626 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-49856262016-08-22 The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes Estandarte, Ana Katrina Botchway, Stanley Lynch, Christophe Yusuf, Mohammed Robinson, Ian Sci Rep Article Chromatin undergoes dramatic condensation and decondensation as cells transition between the different phases of the cell cycle. The organization of chromatin in chromosomes is still one of the key challenges in structural biology. Fluorescence lifetime imaging (FLIM), a technique which utilizes a fluorophore’s fluorescence lifetime to probe changes in its environment, was used to investigate variations in chromatin compaction in fixed human chromosomes. Fixed human metaphase and interphase chromosomes were labeled with the DNA minor groove binder, DAPI, followed by measurement and imaging of the fluorescence lifetime using multiphoton excitation. DAPI lifetime variations in metaphase chromosome spreads allowed mapping of the differentially compacted regions of chromatin along the length of the chromosomes. The heteromorphic regions of chromosomes 1, 9, 15, 16, and Y, which consist of highly condensed constitutive heterochromatin, showed statistically significant shorter DAPI lifetime values than the rest of the chromosomes. Differences in the DAPI lifetimes for the heteromorphic regions suggest differences in the structures of these regions. DAPI lifetime variations across interphase nuclei showed variation in chromatin compaction in interphase and the formation of chromosome territories. The successful probing of differences in chromatin compaction suggests that FLIM has enormous potential for application in structural and diagnostic studies. Nature Publishing Group 2016-08-16 /pmc/articles/PMC4985626/ /pubmed/27526631 http://dx.doi.org/10.1038/srep31417 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Estandarte, Ana Katrina Botchway, Stanley Lynch, Christophe Yusuf, Mohammed Robinson, Ian The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes |
title | The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes |
title_full | The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes |
title_fullStr | The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes |
title_full_unstemmed | The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes |
title_short | The use of DAPI fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes |
title_sort | use of dapi fluorescence lifetime imaging for investigating chromatin condensation in human chromosomes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4985626/ https://www.ncbi.nlm.nih.gov/pubmed/27526631 http://dx.doi.org/10.1038/srep31417 |
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