title: Light-sheet Bayesian microscopy enables deep-cell super-resolution imaging of heterochromatin in live human embryonic stem cells.
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.
  
  
  CONCLUSIONS
  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.
authors: Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors: Ying S
authors: YS
authors: Hu
authors: Laboratory of Genetics, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors: Quan
authors: Q
authors: Zhu
authors: Laboratory of Genetics, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors: Keri
authors: K
authors: Elkins
authors: Department of Bioengineering, University of California San Diego, La Jolla, CA 92093, USA.
authors: Kevin
authors: K
authors: Tse
authors: Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors: Yu
authors: Y
authors: Li
authors: Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors: James A J
authors: JA
authors: Fitzpatrick
authors: Laboratory of Genetics, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors: Inder M
authors: IM
authors: Verma
authors: Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors: Hu
authors: H
authors: Cang
keywords: Bayesian; Heterochromatin; Human embryonic stem cell; Light sheet; Super-resolution imaging