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

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

doi:10.1186/2192-2853-2-7

Title: Light-sheet Bayesian microscopy enables deep-cell super-resolution imaging of heterochromatin in live human embryonic stem cells

Journal Article · Wed Nov 20 00:00:00 EST 2013 · Optical Nanoscopy

DOI:https://doi.org/10.1186/2192-2853-2-7· OSTI ID:1629936

Hu, Ying S. ^[1]; Zhu, Quan ^[2]; Elkins, Keri ^[2]; Tse, Kevin ^[3]; Li, Yu ^[1]; Fitzpatrick, James A. J. ^[1]; Verma, Inder M. ^[2]; Cang, Hu ^[1]

Salk Institute for Biological Studies, La Jolla, CA (United States). Waitt Advanced Biophotonics Center
Salk Institute for Biological Studies, La Jolla, CA (United States). Lab. of Genetics
Univ. of California, San Diego, La Jolla, CA (United States). Dept. of Bioengineering

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 lightsheet 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.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). National Energy Research Scientific Computing Center (NERSC)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1629936

Journal Information:: Optical Nanoscopy, Vol. 2, Issue 1; ISSN 2192-2853

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Super-resolution imaging
Light sheet
Bayesian
Heterochromatin
Human embryonic stem cell

Title: Light-sheet Bayesian microscopy enables deep-cell super-resolution imaging of heterochromatin in live human embryonic stem cells

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