Enhanced FIB-SEM systems for large-volume 3D imaging

ELife
C Shan XuHarald F Hess

Abstract

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 > 106µm3. 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,Drosophilabrain, andChlamydomonas reinhardtiito illustrate the power of this novel high-resolution technique to address questions in both connectomics and cell biology.

References

Jan 1, 1996·Journal of Structural Biology·J R KremerJ R McIntosh
Sep 27, 2000·Current Topics in Developmental Biology·A M PrebleS K Dutcher
May 5, 2001·Annual Review of Plant Physiology and Plant Molecular Biology·Elizabeth H Harris
Mar 21, 2008·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Graham KnottBen Lich
Nov 5, 2011·Science·Jeff W Lichtman, Winfried Denk
Nov 29, 2011·Current Opinion in Neurobiology·Kevin L Briggman, Davi D Bock
Feb 23, 2012·Nature Reviews. Neuroscience·Winfried DenkMoritz Helmstaedter
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Feb 6, 2013·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Jonas BindingWinfried Denk
Aug 9, 2013·Nature·Shin-ya TakemuraDmitri B Chklovskii
Oct 11, 2013·Journal of Microscopy·K M Boergens, W Denk
Apr 14, 2015·Nature Methods·Shawn Mikula, Winfried Denk
Oct 21, 2015·Proceedings of the National Academy of Sciences of the United States of America·Shin-Ya TakemuraLouis K Scheffer
Oct 21, 2015·The Journal of Cell Biology·Jieyan V ChenTimothy L Megraw
Oct 30, 2015·Nature Methods·Kedar Narayan, Sriram Subramaniam
Jun 23, 2016·Journal of Neurogenetics·Ian A Meinertzhagen
Nov 4, 2016·Biology of the Cell·Benjamin Titze, Christel Genoud
Jun 1, 2017·Proceedings of the National Academy of Sciences of the United States of America·Yumei WuPietro De Camilli

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Citations

May 24, 2018·Journal of Neurogenetics·Ian A Meinertzhagen
Jul 19, 2017·ELife·Shin-Ya TakemuraLouis K Scheffer
Sep 25, 2018·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Manja Luckner, Gerhard Wanner
Jan 10, 2019·ELife·Kazunori ShinomiyaIan A Meinertzhagen
Apr 25, 2018·Emerging Microbes & Infections·Jun LiuXiankun Zeng
Aug 1, 2019·Biology Open·Thomas DelgadoPamela J Yao
Sep 7, 2019·Acta Neuropathologica Communications·Whitney C McFaddenAndrew T McKenzie
Sep 26, 2019·Proceedings of the National Academy of Sciences of the United States of America·Satoshi OkadaKen Takai
Dec 14, 2019·The Journal of Cell Biology·Mohammad RahmanOrna Cohen-Fix
Jun 9, 2020·PLoS Pathogens·Rachel M RudlaffJeffrey D Dvorin
May 6, 2020·The Journal of Cell Biology·Andreas Prokop
May 24, 2018·Histochemistry and Cell Biology·Manja Luckner, Gerhard Wanner
Nov 30, 2018·Frontiers in Neural Circuits·Yoshiyuki KubotaYasuo Kawaguchi
Mar 23, 2019·Frontiers in Genetics·Sarah C Hughes, Andrew J Simmonds
Sep 12, 2019·Frontiers in Neuroscience·Christine SaleebaSimon McMullan
Jun 23, 2019·The Journal of Cell Biology·Chi-Lun ChangJennifer Lippincott-Schwartz
Mar 30, 2020·Bioinformatics·Jose-Jesus FernandezMaria-Rosario Fernandez-Fernandez
Jun 1, 2017·Proceedings of the National Academy of Sciences of the United States of America·Yumei WuPietro De Camilli
Nov 30, 2018·Frontiers in Neural Circuits·Ting ZhaoStephen M Plaza
May 15, 2020·Proceedings of the National Academy of Sciences of the United States of America·José G García-CerdánKrishna K Niyogi
May 23, 2018·The Journal of Experimental Biology·Ian A Meinertzhagen
Jan 9, 2019·Frontiers in Neuroanatomy·Anna Lena Eberle, Dirk Zeidler
Sep 16, 2020·Nature Neuroscience·Aaron T KuanWei-Chung Allen Lee
Oct 21, 2020·Nature·Esther Landhuis
Sep 4, 2020·ELife·Louis K SchefferStephen M Plaza
Oct 30, 2020·Frontiers in Bioengineering and Biotechnology·Mootaz M SalmanTom Kirchhausen
Sep 18, 2020·The Journal of Biological Chemistry·Peter O BayguinovJames A J Fitzpatrick
Dec 17, 2020·The Journal of Cell Biology·Andreas MüllerMichele Solimena
Dec 31, 2020·STAR Protocols·Georg KislingerMartina Schifferer
Sep 18, 2020·IScience·Pamela J YaoDimitrios Kapogiannis
Oct 22, 2019·Trends in Cell Biology·Janine McCaughey, David J Stephens

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Methods Mentioned

BETA
electron microscopy
X-ray
fluorescence microscopy
electron scattering
transmission electron microscopy
scanning electron microscopy
chemical fixation

Software Mentioned

SIFT
Fiji
IMOD
SEM
SIMION
FLYEM
InLens
LabVIEW
FIB

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