Quantifying nuclear wide chromatin compaction by phasor analysis of histone Förster resonance energy transfer (FRET) in frequency domain fluorescence lifetime imaging microscopy (FLIM) data
The nanometer spacing between nucleosomes throughout global chromatin organisation modulates local DNA template access, and through continuous dynamic rearrangements, regulates genome function [1]. However, given that nucleosome packaging occurs on a spatial scale well below the diffraction limit, r...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | Data in Brief |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S235234092030295X |
| _version_ | 1819076644184260608 |
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| author | Zhen Liang Jieqiong Lou Lorenzo Scipioni Enrico Gratton Elizabeth Hinde |
| author_facet | Zhen Liang Jieqiong Lou Lorenzo Scipioni Enrico Gratton Elizabeth Hinde |
| author_sort | Zhen Liang |
| collection | DOAJ |
| description | The nanometer spacing between nucleosomes throughout global chromatin organisation modulates local DNA template access, and through continuous dynamic rearrangements, regulates genome function [1]. However, given that nucleosome packaging occurs on a spatial scale well below the diffraction limit, real time observation of chromatin structure in live cells by optical microscopy has proved technically difficult, despite recent advances in live cell super resolution imaging [2]. One alternative solution to quantify chromatin structure in a living cell at the level of nucleosome proximity is to measure and spatially map Förster resonance energy transfer (FRET) between fluorescently labelled histones – the core protein of a nucleosome [3]. In recent work we established that the phasor approach to fluorescence lifetime imaging microscopy (FLIM) is a robust method for the detection of histone FRET which can quantify nuclear wide chromatin compaction in the presence of cellular autofluorescence [4]. Here we share FLIM data recording histone FRET in live cells co-expressing H2B-eGFP and H2B-mCherry. The data was acquired in the frequency domain [5] and processed by the phasor approach to lifetime analysis [6]. The data can be valuable to researchers interested in using the histone FRET assay since it highlights the impact of cellular autofluorescence and acceptor-donor ratio on quantifying chromatin compaction. The data is related to the research article “Phasor histone FLIM-FRET microscopy quantifies spatiotemporal rearrangement of chromatin architecture during the DNA damage response” [4]. |
| first_indexed | 2024-12-21T18:44:35Z |
| format | Article |
| id | doaj.art-e7a62209a8504884bab49b87bdc8c221 |
| institution | Directory Open Access Journal |
| issn | 2352-3409 |
| language | English |
| last_indexed | 2024-12-21T18:44:35Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Data in Brief |
| spelling | doaj.art-e7a62209a8504884bab49b87bdc8c2212022-12-21T18:53:55ZengElsevierData in Brief2352-34092020-06-0130105401Quantifying nuclear wide chromatin compaction by phasor analysis of histone Förster resonance energy transfer (FRET) in frequency domain fluorescence lifetime imaging microscopy (FLIM) dataZhen Liang0Jieqiong Lou1Lorenzo Scipioni2Enrico Gratton3Elizabeth Hinde4School of Physics, University of Melbourne, Australia; Department of Biochemistry and Molecular Biology, Bio21 Institute, University of Melbourne, AustraliaSchool of Physics, University of Melbourne, Australia; Department of Biochemistry and Molecular Biology, Bio21 Institute, University of Melbourne, AustraliaDepartment of Biomedical Engineering, Laboratory for Fluorescence Dynamics, University of California, Irvine, United StatesDepartment of Biomedical Engineering, Laboratory for Fluorescence Dynamics, University of California, Irvine, United StatesSchool of Physics, University of Melbourne, Australia; Department of Biochemistry and Molecular Biology, Bio21 Institute, University of Melbourne, Australia; Corresponding author at: School of Physics, University of Melbourne, Australia.The nanometer spacing between nucleosomes throughout global chromatin organisation modulates local DNA template access, and through continuous dynamic rearrangements, regulates genome function [1]. However, given that nucleosome packaging occurs on a spatial scale well below the diffraction limit, real time observation of chromatin structure in live cells by optical microscopy has proved technically difficult, despite recent advances in live cell super resolution imaging [2]. One alternative solution to quantify chromatin structure in a living cell at the level of nucleosome proximity is to measure and spatially map Förster resonance energy transfer (FRET) between fluorescently labelled histones – the core protein of a nucleosome [3]. In recent work we established that the phasor approach to fluorescence lifetime imaging microscopy (FLIM) is a robust method for the detection of histone FRET which can quantify nuclear wide chromatin compaction in the presence of cellular autofluorescence [4]. Here we share FLIM data recording histone FRET in live cells co-expressing H2B-eGFP and H2B-mCherry. The data was acquired in the frequency domain [5] and processed by the phasor approach to lifetime analysis [6]. The data can be valuable to researchers interested in using the histone FRET assay since it highlights the impact of cellular autofluorescence and acceptor-donor ratio on quantifying chromatin compaction. The data is related to the research article “Phasor histone FLIM-FRET microscopy quantifies spatiotemporal rearrangement of chromatin architecture during the DNA damage response” [4].http://www.sciencedirect.com/science/article/pii/S235234092030295XFörster resonance energy transferFluorescence lifetime imaging microscopyPhasor analysisHistoneChromatin compactionNuclear architecture |
| spellingShingle | Zhen Liang Jieqiong Lou Lorenzo Scipioni Enrico Gratton Elizabeth Hinde Quantifying nuclear wide chromatin compaction by phasor analysis of histone Förster resonance energy transfer (FRET) in frequency domain fluorescence lifetime imaging microscopy (FLIM) data Data in Brief Förster resonance energy transfer Fluorescence lifetime imaging microscopy Phasor analysis Histone Chromatin compaction Nuclear architecture |
| title | Quantifying nuclear wide chromatin compaction by phasor analysis of histone Förster resonance energy transfer (FRET) in frequency domain fluorescence lifetime imaging microscopy (FLIM) data |
| title_full | Quantifying nuclear wide chromatin compaction by phasor analysis of histone Förster resonance energy transfer (FRET) in frequency domain fluorescence lifetime imaging microscopy (FLIM) data |
| title_fullStr | Quantifying nuclear wide chromatin compaction by phasor analysis of histone Förster resonance energy transfer (FRET) in frequency domain fluorescence lifetime imaging microscopy (FLIM) data |
| title_full_unstemmed | Quantifying nuclear wide chromatin compaction by phasor analysis of histone Förster resonance energy transfer (FRET) in frequency domain fluorescence lifetime imaging microscopy (FLIM) data |
| title_short | Quantifying nuclear wide chromatin compaction by phasor analysis of histone Förster resonance energy transfer (FRET) in frequency domain fluorescence lifetime imaging microscopy (FLIM) data |
| title_sort | quantifying nuclear wide chromatin compaction by phasor analysis of histone forster resonance energy transfer fret in frequency domain fluorescence lifetime imaging microscopy flim data |
| topic | Förster resonance energy transfer Fluorescence lifetime imaging microscopy Phasor analysis Histone Chromatin compaction Nuclear architecture |
| url | http://www.sciencedirect.com/science/article/pii/S235234092030295X |
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