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Title: 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 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 > 10 6 ?m 3 . 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.
Authors:
ORCiD logo [1] ;  [1] ;  [2] ;  [3] ;  [4] ;  [3] ;  [5] ; ORCiD logo [5] ;  [6] ;  [1]
  1. Howard Hughes Medical Institute, Ashburn, VA (United States)
  2. Howard Hughes Medical Institute, Ashburn, VA (United States); Dalhousie Univ., Halifax (Canada)
  3. Univ. of California, Berkeley, CA (United States)
  4. Univ. of California, Berkeley, CA (United States); Univ. of Texas, Austin, TX (United States)
  5. Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  6. Univ. of North Carolina, Chapel Hill, NC (United States)
Publication Date:
Grant/Contract Number:
AC02-05CH11231; Office of Science, Basic Energy Sciences, Chemical Sciences, Geosciences, and Biosciences Division - SISGRKN
Type:
Published Article
Journal Name:
eLife
Additional Journal Information:
Journal Volume: 6; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Research Org:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1363984
Alternate Identifier(s):
OSTI ID: 1363985; OSTI ID: 1379848

Xu, C. Shan, Hayworth, Kenneth J., Lu, Zhiyuan, Grob, Patricia, Hassan, Ahmed M., Garcia-Cerdan, Jose G., Niyogi, Krishna K., Nogales, Eva, Weinberg, Richard J., and Hess, Harald F.. Enhanced FIB-SEM systems for large-volume 3D imaging. United States: N. p., Web. doi:10.7554/eLife.25916.
Xu, C. Shan, Hayworth, Kenneth J., Lu, Zhiyuan, Grob, Patricia, Hassan, Ahmed M., Garcia-Cerdan, Jose G., Niyogi, Krishna K., Nogales, Eva, Weinberg, Richard J., & Hess, Harald F.. Enhanced FIB-SEM systems for large-volume 3D imaging. United States. doi:10.7554/eLife.25916.
Xu, C. Shan, Hayworth, Kenneth J., Lu, Zhiyuan, Grob, Patricia, Hassan, Ahmed M., Garcia-Cerdan, Jose G., Niyogi, Krishna K., Nogales, Eva, Weinberg, Richard J., and Hess, Harald F.. 2017. "Enhanced FIB-SEM systems for large-volume 3D imaging". United States. doi:10.7554/eLife.25916.
@article{osti_1363984,
title = {Enhanced FIB-SEM systems for large-volume 3D imaging},
author = {Xu, C. Shan and Hayworth, Kenneth J. and Lu, Zhiyuan and Grob, Patricia and Hassan, Ahmed M. and Garcia-Cerdan, Jose G. and Niyogi, Krishna K. and Nogales, Eva and Weinberg, Richard J. and Hess, Harald F.},
abstractNote = {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 > 10 6 ?m 3 . 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.},
doi = {10.7554/eLife.25916},
journal = {eLife},
number = ,
volume = 6,
place = {United States},
year = {2017},
month = {5}
}

Works referenced in this record:

Serial Block-Face Scanning Electron Microscopy to Reconstruct Three-Dimensional Tissue Nanostructure
journal, October 2004