Enhanced FIB-SEM systems for large-volume 3D imaging
Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) can automatically generate 3D images with superior z-axis resolution, yielding data that needs minimal image registration and related post-processing. Obstacles blocking wider adoption of FIB-SEM include slow imaging speed and lack of long-term...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2017-05-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/25916 |
| _version_ | 1811253350826508288 |
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| author | C Shan Xu Kenneth J Hayworth Zhiyuan Lu Patricia Grob Ahmed M Hassan José G García-Cerdán Krishna K Niyogi Eva Nogales Richard J Weinberg Harald F Hess |
| author_facet | C Shan Xu Kenneth J Hayworth Zhiyuan Lu Patricia Grob Ahmed M Hassan José G García-Cerdán Krishna K Niyogi Eva Nogales Richard J Weinberg Harald F Hess |
| author_sort | C Shan Xu |
| collection | DOAJ |
| description | Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) can automatically generate 3D images with superior z-axis resolution, yielding data that needs minimal image registration and related post-processing. Obstacles blocking wider adoption of FIB-SEM include slow imaging speed and lack of long-term system stability, which caps the maximum possible acquisition volume. Here, we present techniques that accelerate image acquisition while greatly improving FIB-SEM reliability, allowing the system to operate for months and generating continuously imaged volumes > 106 µm3. These volumes are large enough for connectomics, where the excellent z resolution can help in tracing of small neuronal processes and accelerate the tedious and time-consuming human proofreading effort. Even higher resolution can be achieved on smaller volumes. We present example data sets from mammalian neural tissue, Drosophila brain, and Chlamydomonas reinhardtii to illustrate the power of this novel high-resolution technique to address questions in both connectomics and cell biology. |
| first_indexed | 2024-04-12T16:49:36Z |
| format | Article |
| id | doaj.art-1ba7c7928d214f728c18bc556afa0243 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:49:36Z |
| publishDate | 2017-05-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-1ba7c7928d214f728c18bc556afa02432022-12-22T03:24:27ZengeLife Sciences Publications LtdeLife2050-084X2017-05-01610.7554/eLife.25916Enhanced FIB-SEM systems for large-volume 3D imagingC Shan Xu0https://orcid.org/0000-0002-8564-7836Kenneth J Hayworth1Zhiyuan Lu2Patricia Grob3Ahmed M Hassan4José G García-Cerdán5Krishna K Niyogi6Eva Nogales7https://orcid.org/0000-0001-9816-3681Richard J Weinberg8Harald F Hess9Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United StatesJanelia Research Campus, Howard Hughes Medical Institute, Ashburn, United StatesJanelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States; Department of Psychology and Neuroscience, Dalhousie University, Halifax, CanadaHoward Hughes Medical Institute, Molecular and Cell Biology Department, University of California, Berkeley, United StatesHoward Hughes Medical Institute, Molecular and Cell Biology Department, University of California, Berkeley, United StatesHoward Hughes Medical Institute, Plant and Microbial Biology Department, University of California, Berkeley, United StatesHoward Hughes Medical Institute, Plant and Microbial Biology Department, University of California, Berkeley, United States; Lawrence Berkeley National Laboratory, Berkeley, United StatesHoward Hughes Medical Institute, Molecular and Cell Biology Department, University of California, Berkeley, United States; Lawrence Berkeley National Laboratory, Berkeley, United StatesDepartment of Cell Biology and Physiology, University of North Carolina, North Carolina, United StatesJanelia Research Campus, Howard Hughes Medical Institute, Ashburn, United StatesFocused Ion Beam Scanning Electron Microscopy (FIB-SEM) can automatically generate 3D images with superior z-axis resolution, yielding data that needs minimal image registration and related post-processing. Obstacles blocking wider adoption of FIB-SEM include slow imaging speed and lack of long-term system stability, which caps the maximum possible acquisition volume. Here, we present techniques that accelerate image acquisition while greatly improving FIB-SEM reliability, allowing the system to operate for months and generating continuously imaged volumes > 106 µm3. These volumes are large enough for connectomics, where the excellent z resolution can help in tracing of small neuronal processes and accelerate the tedious and time-consuming human proofreading effort. Even higher resolution can be achieved on smaller volumes. We present example data sets from mammalian neural tissue, Drosophila brain, and Chlamydomonas reinhardtii to illustrate the power of this novel high-resolution technique to address questions in both connectomics and cell biology.https://elifesciences.org/articles/25916connectomics3D cellular structureelectron microscopyimagingFIB |
| spellingShingle | C Shan Xu Kenneth J Hayworth Zhiyuan Lu Patricia Grob Ahmed M Hassan José G García-Cerdán Krishna K Niyogi Eva Nogales Richard J Weinberg Harald F Hess Enhanced FIB-SEM systems for large-volume 3D imaging eLife connectomics 3D cellular structure electron microscopy imaging FIB |
| title | Enhanced FIB-SEM systems for large-volume 3D imaging |
| title_full | Enhanced FIB-SEM systems for large-volume 3D imaging |
| title_fullStr | Enhanced FIB-SEM systems for large-volume 3D imaging |
| title_full_unstemmed | Enhanced FIB-SEM systems for large-volume 3D imaging |
| title_short | Enhanced FIB-SEM systems for large-volume 3D imaging |
| title_sort | enhanced fib sem systems for large volume 3d imaging |
| topic | connectomics 3D cellular structure electron microscopy imaging FIB |
| url | https://elifesciences.org/articles/25916 |
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