Gas cluster ion beam SEM for imaging of large tissue samples with 10 nm isotropic resolution
We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be...
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| Published in | Nature methods Vol. 17; no. 1; pp. 68 - 71 |
|---|---|
| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
New York
Nature Publishing Group US
01.01.2020
Nature Publishing Group |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1548-7091 1548-7105 1548-7105 |
| DOI | 10.1038/s41592-019-0641-2 |
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| Abstract | We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to span the sectioned volume. Incorporating gas cluster ion beam SEM into existing single-beam and multibeam SEM workflows should be straightforward, increasing reliability while improving
z
resolution by a factor of three or more.
An alternative to focused ion beam scanning electron microscopy (FIB-SEM), gas cluster ion beam scanning electron microscopy (GCIB-SEM) is compatible with large tissue samples while achieving similar isotropic resolution. |
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| AbstractList | We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to span the sectioned volume. Incorporating gas cluster ion beam SEM into existing single-beam and multibeam SEM workflows should be straightforward, increasing reliability while improving z resolution by a factor of three or more. We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to span the sectioned volume. Incorporating gas cluster ion beam SEM into existing single-beam and multibeam SEM workflows should be straightforward, increasing reliability while improving z resolution by a factor of three or more. An alternative to focused ion beam scanning electron microscopy (FIB-SEM), gas cluster ion beam scanning electron microscopy (GCIB-SEM) is compatible with large tissue samples while achieving similar isotropic resolution. We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to span the sectioned volume. Incorporating gas cluster ion beam SEM into existing single-beam and multibeam SEM workflows should be straightforward, increasing reliability while improving z resolution by a factor of three or more. An alternative to focused ion beam scanning electron microscopy (FIB-SEM), gas cluster ion beam scanning electron microscopy (GCIB-SEM) is compatible with large tissue samples while achieving similar isotropic resolution. We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to span the sectioned volume. Incorporating gas cluster ion beam SEM into existing single-beam and multibeam SEM workflows should be straightforward, increasing reliability while improving z resolution by a factor of three or more.We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This three-dimensional electron microscopy technique acquires datasets with <10 nm isotropic resolution of each section, and these can then be stitched together to span the sectioned volume. Incorporating gas cluster ion beam SEM into existing single-beam and multibeam SEM workflows should be straightforward, increasing reliability while improving z resolution by a factor of three or more. |
| Audience | Academic |
| Author | Lu, Zhiyuan Xu, C. Shan Peale, David Januszewski, Michał Hess, Harald F. Hayworth, Kenneth J. Knott, Graham W. |
| Author_xml | – sequence: 1 givenname: Kenneth J. orcidid: 0000-0002-6467-7024 surname: Hayworth fullname: Hayworth, Kenneth J. email: hayworthk@janelia.hhmi.org organization: HHMI Janelia Research Campus – sequence: 2 givenname: David surname: Peale fullname: Peale, David organization: HHMI Janelia Research Campus – sequence: 3 givenname: Michał orcidid: 0000-0002-3480-2744 surname: Januszewski fullname: Januszewski, Michał organization: Google Research – sequence: 4 givenname: Graham W. orcidid: 0000-0002-2956-9052 surname: Knott fullname: Knott, Graham W. organization: EPFL – sequence: 5 givenname: Zhiyuan surname: Lu fullname: Lu, Zhiyuan organization: Dalhousie University – sequence: 6 givenname: C. Shan surname: Xu fullname: Xu, C. Shan organization: HHMI Janelia Research Campus – sequence: 7 givenname: Harald F. orcidid: 0000-0003-3000-1533 surname: Hess fullname: Hess, Harald F. organization: HHMI Janelia Research Campus |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31740820$$D View this record in MEDLINE/PubMed |
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| Snippet | We demonstrate gas cluster ion beam scanning electron microscopy (SEM), in which wide-area ion milling is performed on a series of thick tissue sections. This... |
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| SubjectTerms | 631/1647/328/1649 631/1647/334/1582/715 631/1647/334/1874/345 631/378 631/378/2597 Animals Bioinformatics Biological Microscopy Biological Techniques Biomedical and Life Sciences Biomedical Engineering/Biotechnology Brain - ultrastructure Brief Communication Cerebral Cortex - ultrastructure Clusters Drosophila melanogaster Electron microscopy Image Processing, Computer-Assisted - methods Imaging, Three-Dimensional - methods Ion beams Life Sciences Male Mice Mice, Inbred C57BL Microscopy, Electron, Scanning - methods Proteomics Reliability aspects Scanning electron microscopy Tissue Fixation |
| Title | Gas cluster ion beam SEM for imaging of large tissue samples with 10 nm isotropic resolution |
| URI | https://link.springer.com/article/10.1038/s41592-019-0641-2 https://www.ncbi.nlm.nih.gov/pubmed/31740820 https://www.proquest.com/docview/2333915591 https://www.proquest.com/docview/2315970812 |
| Volume | 17 |
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