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Volumetric interferometric lattice light-sheet imaging
Live-cell imaging with high spatiotemporal resolution and high detection sensitivity facilitates the study of the dynamics of cellular structure and function. However, extracting high-resolution 4D (3D space plus time) information from live cells remains challenging, because current methods are slow...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8595582/ https://www.ncbi.nlm.nih.gov/pubmed/34635835 http://dx.doi.org/10.1038/s41587-021-01042-y |
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author | Cao, Bin Coelho, Simao Li, Jieru Wang, Guanshi Pertsinidis, Alexandros |
author_facet | Cao, Bin Coelho, Simao Li, Jieru Wang, Guanshi Pertsinidis, Alexandros |
author_sort | Cao, Bin |
collection | PubMed |
description | Live-cell imaging with high spatiotemporal resolution and high detection sensitivity facilitates the study of the dynamics of cellular structure and function. However, extracting high-resolution 4D (3D space plus time) information from live cells remains challenging, because current methods are slow, require high peak excitation intensities or suffer from high out-of-focus background. Here we present 3D interferometric lattice light-sheet (3D-iLLS) imaging, a technique that requires low excitation light levels and provides high background suppression and substantially improved volumetric resolution by combining 4Pi interferometry with selective plane illumination. We demonstrate that 3D-iLLS has an axial resolution and single-particle localization precision of 100 nm (FWHM) and <10 nm (1σ), respectively. We illustrate the performance of 3D-iLLS in a range of systems: single messenger RNA molecules, nanoscale assemblies of transcription regulators in the nucleus, the microtubule cytoskeleton, and mitochondria organelles. The enhanced 4D resolution and increased signal-to-noise ratio (SNR) of 3D-iLLS will facilitate the analysis of biological processes at the sub-cellular level. |
format | Online Article Text |
id | pubmed-8595582 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
record_format | MEDLINE/PubMed |
spelling | pubmed-85955822022-04-11 Volumetric interferometric lattice light-sheet imaging Cao, Bin Coelho, Simao Li, Jieru Wang, Guanshi Pertsinidis, Alexandros Nat Biotechnol Article Live-cell imaging with high spatiotemporal resolution and high detection sensitivity facilitates the study of the dynamics of cellular structure and function. However, extracting high-resolution 4D (3D space plus time) information from live cells remains challenging, because current methods are slow, require high peak excitation intensities or suffer from high out-of-focus background. Here we present 3D interferometric lattice light-sheet (3D-iLLS) imaging, a technique that requires low excitation light levels and provides high background suppression and substantially improved volumetric resolution by combining 4Pi interferometry with selective plane illumination. We demonstrate that 3D-iLLS has an axial resolution and single-particle localization precision of 100 nm (FWHM) and <10 nm (1σ), respectively. We illustrate the performance of 3D-iLLS in a range of systems: single messenger RNA molecules, nanoscale assemblies of transcription regulators in the nucleus, the microtubule cytoskeleton, and mitochondria organelles. The enhanced 4D resolution and increased signal-to-noise ratio (SNR) of 3D-iLLS will facilitate the analysis of biological processes at the sub-cellular level. 2021-10-11 2021-11 /pmc/articles/PMC8595582/ /pubmed/34635835 http://dx.doi.org/10.1038/s41587-021-01042-y Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: https://www.springernature.com/gp/open-research/policies/accepted-manuscript-terms |
spellingShingle | Article Cao, Bin Coelho, Simao Li, Jieru Wang, Guanshi Pertsinidis, Alexandros Volumetric interferometric lattice light-sheet imaging |
title | Volumetric interferometric lattice light-sheet imaging |
title_full | Volumetric interferometric lattice light-sheet imaging |
title_fullStr | Volumetric interferometric lattice light-sheet imaging |
title_full_unstemmed | Volumetric interferometric lattice light-sheet imaging |
title_short | Volumetric interferometric lattice light-sheet imaging |
title_sort | volumetric interferometric lattice light-sheet imaging |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8595582/ https://www.ncbi.nlm.nih.gov/pubmed/34635835 http://dx.doi.org/10.1038/s41587-021-01042-y |
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