Light-sheet Bayesian microscopy enables deep-cell super-resolution imaging of heterochromatin in live human embryonic stem cells
Background Heterochromatin in the nucleus of human embryonic cells plays an important role in the epigenetic regulation of gene expression. The architecture of heterochromatin and its dynamic organization remain elusive because of the lack of fast and high-resolution deep-cell imaging tools. We enab...
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| Vydáno v: | Optical nanoscopy Ročník 2; číslo 1; s. 1 - 12 |
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| Médium: | Journal Article |
| Jazyk: | angličtina |
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Berlin/Heidelberg
Springer Berlin Heidelberg
20.11.2013
Springer Nature B.V |
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| ISSN: | 2192-2853 |
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| Abstract | Background
Heterochromatin in the nucleus of human embryonic cells plays an important role in the epigenetic regulation of gene expression. The architecture of heterochromatin and its dynamic organization remain elusive because of the lack of fast and high-resolution deep-cell imaging tools. We enable this task by advancing instrumental and algorithmic implementation of the localization-based super-resolution technique.
Results
We present light-sheet Bayesian super-resolution microscopy (LSBM). We adapt light-sheet illumination for super-resolution imaging by using a novel prism-coupled condenser design to illuminate a thin slice of the nucleus with high signal-to-noise ratio. Coupled with a Bayesian algorithm that resolves overlapping fluorophores from high-density areas, we show, for the first time, nanoscopic features of the heterochromatin structure in both fixed and live human embryonic stem cells. The enhanced temporal resolution allows capturing the dynamic change of heterochromatin with a lateral resolution of 50-60 nm on a time scale of 2.3 s.
Conclusion
Light-sheet Bayesian microscopy opens up broad new possibilities of probing nanometer-scale nuclear structures and real-time sub-cellular processes and other previously difficult-to-access intracellular regions of living cells at the single-molecule, and single cell level. |
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| AbstractList | Heterochromatin in the nucleus of human embryonic cells plays an important role in the epigenetic regulation of gene expression. The architecture of heterochromatin and its dynamic organization remain elusive because of the lack of fast and high-resolution deep-cell imaging tools. We enable this task by advancing instrumental and algorithmic implementation of the localization-based super-resolution technique. We present light-sheet Bayesian super-resolution microscopy (LSBM). We adapt light-sheet illumination for super-resolution imaging by using a novel prism-coupled condenser design to illuminate a thin slice of the nucleus with high signal-to-noise ratio. Coupled with a Bayesian algorithm that resolves overlapping fluorophores from high-density areas, we show, for the first time, nanoscopic features of the heterochromatin structure in both fixed and live human embryonic stem cells. The enhanced temporal resolution allows capturing the dynamic change of heterochromatin with a lateral resolution of 50-60 nm on a time scale of 2.3 s. Light-sheet Bayesian microscopy opens up broad new possibilities of probing nanometer-scale nuclear structures and real-time sub-cellular processes and other previously difficult-to-access intracellular regions of living cells at the single-molecule, and single cell level. Background: Heterochromatin in the nucleus of human embryonic cells plays an important role in the epigenetic regulation of gene expression. The architecture of heterochromatin and its dynamic organization remain elusive because of the lack of fast and high-resolution deep-cell imaging tools. We enable this task by advancing instrumental and algorithmic implementation of the localization-based super-resolution technique. Results: We present light-sheet Bayesian super-resolution microscopy (LSBM). We adapt light-sheet illumination for super-resolution imaging by using a novel prism-coupled condenser design to illuminate a thin slice of the nucleus with high signal-to-noise ratio. Coupled with a Bayesian algorithm that resolves overlapping fluorophores from high-density areas, we show, for the first time, nanoscopic features of the heterochromatin structure in both fixed and live human embryonic stem cells. The enhanced temporal resolution allows capturing the dynamic change of heterochromatin with a lateral resolution of 50-60 nm on a time scale of 2.3 s. Conclusion: Light-sheet Bayesian microscopy opens up broad new possibilities of probing nanometer-scale nuclear structures and real-time sub-cellular processes and other previously difficult-to-access intracellular regions of living cells at the single-molecule, and single cell level. Background Heterochromatin in the nucleus of human embryonic cells plays an important role in the epigenetic regulation of gene expression. The architecture of heterochromatin and its dynamic organization remain elusive because of the lack of fast and high-resolution deep-cell imaging tools. We enable this task by advancing instrumental and algorithmic implementation of the localization-based super-resolution technique. Results We present light-sheet Bayesian super-resolution microscopy (LSBM). We adapt light-sheet illumination for super-resolution imaging by using a novel prism-coupled condenser design to illuminate a thin slice of the nucleus with high signal-to-noise ratio. Coupled with a Bayesian algorithm that resolves overlapping fluorophores from high-density areas, we show, for the first time, nanoscopic features of the heterochromatin structure in both fixed and live human embryonic stem cells. The enhanced temporal resolution allows capturing the dynamic change of heterochromatin with a lateral resolution of 50-60 nm on a time scale of 2.3 s. Conclusion Light-sheet Bayesian microscopy opens up broad new possibilities of probing nanometer-scale nuclear structures and real-time sub-cellular processes and other previously difficult-to-access intracellular regions of living cells at the single-molecule, and single cell level. |
| Author | Fitzpatrick, James A J Li, Yu Elkins, Keri Verma, Inder M Cang, Hu Hu, Ying S Tse, Kevin Zhu, Quan |
| AuthorAffiliation | 2 Laboratory of Genetics, Salk Institute for Biological Studies, La Jolla, CA 92037, USA 3 Department of Bioengineering, University of California San Diego, La Jolla, CA 92093, USA 1 Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037, USA |
| AuthorAffiliation_xml | – name: 2 Laboratory of Genetics, Salk Institute for Biological Studies, La Jolla, CA 92037, USA – name: 3 Department of Bioengineering, University of California San Diego, La Jolla, CA 92093, USA – name: 1 Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037, USA |
| Author_xml | – sequence: 1 givenname: Ying S surname: Hu fullname: Hu, Ying S organization: Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies – sequence: 2 givenname: Quan surname: Zhu fullname: Zhu, Quan email: qzhu@salk.edu organization: Laboratory of Genetics, Salk Institute for Biological Studies – sequence: 3 givenname: Keri surname: Elkins fullname: Elkins, Keri organization: Laboratory of Genetics, Salk Institute for Biological Studies, Department of Cellular and Molecular Medicine, University of California San Diego – sequence: 4 givenname: Kevin surname: Tse fullname: Tse, Kevin organization: Department of Bioengineering, University of California San Diego – sequence: 5 givenname: Yu surname: Li fullname: Li, Yu organization: Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies – sequence: 6 givenname: James A J surname: Fitzpatrick fullname: Fitzpatrick, James A J organization: Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies – sequence: 7 givenname: Inder M surname: Verma fullname: Verma, Inder M email: verma@salk.edu organization: Laboratory of Genetics, Salk Institute for Biological Studies – sequence: 8 givenname: Hu surname: Cang fullname: Cang, Hu email: hucang@salk.edu organization: Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies |
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| Keywords | Human embryonic stem cell Heterochromatin Super-resolution imaging Bayesian Light sheet |
| Language | English |
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Heterochromatin in the nucleus of human embryonic cells plays an important role in the epigenetic regulation of gene expression. The architecture of... Heterochromatin in the nucleus of human embryonic cells plays an important role in the epigenetic regulation of gene expression. The architecture of... Background: Heterochromatin in the nucleus of human embryonic cells plays an important role in the epigenetic regulation of gene expression. The architecture... |
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| SubjectTerms | Biochemistry Biological and Medical Physics Biological Microscopy Biological Techniques Biomedical and Life Sciences Biophysics Life Sciences Nanotechnology Original Article |
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| Title | Light-sheet Bayesian microscopy enables deep-cell super-resolution imaging of heterochromatin in live human embryonic stem cells |
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