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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Veröffentlicht in:Optical nanoscopy Jg. 2; H. 1; S. 1 - 12
Hauptverfasser: Hu, Ying S, Zhu, Quan, Elkins, Keri, Tse, Kevin, Li, Yu, Fitzpatrick, James A J, Verma, Inder M, Cang, Hu
Format: Journal Article
Sprache:Englisch
Veröffentlicht: Berlin/Heidelberg Springer Berlin Heidelberg 20.11.2013
Springer Nature B.V
Schlagworte:
Biochemistry
Biological and Medical Physics
Biological Microscopy
Biological Techniques
Biomedical and Life Sciences
Biophysics
Life Sciences
Nanotechnology
Original Article
Human embryonic stem cell
Heterochromatin
Super-resolution imaging
Bayesian
Light sheet
ISSN:2192-2853
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Zusammenfassung: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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Equal contributors
Current address: Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA 92093, USA.
ISSN:2192-2853
DOI:10.1186/2192-2853-2-7