Light-sheet Bayesian microscopy enables deepcell super-resolution imaging of heterochromatin in live human embryonic stem cells

Ying S. Hu, Quan Zhu, Keri Elkins, Kevin Tse, Yu Li, James A.J. Fitzpatrick, Inder M. Verma, Hu Cang

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

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.

Original languageEnglish
Article number7
Pages (from-to)1-12
Number of pages12
JournalOptical Nanoscopy
Volume2
Issue number1
DOIs
StatePublished - 2013
Externally publishedYes

Keywords

  • Bayesian
  • Heterochromatin
  • Human embryonic stem cell
  • Light sheet
  • Super-resolution imaging

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