title: Developing 3D SEM in a broad biological context.
abstract: When electron microscopy (EM) was introduced in the 1930s it gave scientists their first look into the nanoworld of cells. Over the last 80 years EM has vastly increased our understanding of the complex cellular structures that underlie the diverse functions that cells need to maintain life. One drawback that has been difficult to overcome was the inherent lack of volume information, mainly due to the limit on the thickness of sections that could be viewed in a transmission electron microscope (TEM). For many years scientists struggled to achieve three-dimensional (3D) EM using serial section reconstructions, TEM tomography, and scanning EM (SEM) techniques such as freeze-fracture. Although each technique yielded some special information, they required a significant amount of time and specialist expertise to obtain even a very small 3D EM dataset. Almost 20 years ago scientists began to exploit SEMs to image blocks of embedded tissues and perform serial sectioning of these tissues inside the SEM chamber. Using first focused ion beams (FIB) and subsequently robotic ultramicrotomes (serial block-face, SBF-SEM) microscopists were able to collect large volumes of 3D EM information at resolutions that could address many important biological questions, and do so in an efficient manner. We present here some examples of 3D EM taken from the many diverse specimens that have been imaged in our core facility. We propose that the next major step forward will be to efficiently correlate functional information obtained using light microscopy (LM) with 3D EM datasets to more completely investigate the important links between cell structures and their functions.
authors.affiliation: VIB Bio Imaging Core, Gent, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: A
authors.initials: A
authors.lastname: Kremer
authors.affiliation: VIB Bio Imaging Core, Gent, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Lippens
authors.affiliation: VIB Bio Imaging Core, Gent, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Bartunkova
authors.affiliation: VIB Department of Molecular Genetics, Antwerp University, Antwerpen 2020, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Asselbergh
authors.affiliation: IRIBHM, Université Libre de Bruxelles, Brussels, B-1070, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Blanpain
authors.affiliation: Department of Plant Systems Biology, VIB, Ghent, 9052, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052, Ghent, Belgium.|Institute of Science and Technology (IST) Austria, Klosterneuburg, 3400, Austria.
authors.forename: M
authors.initials: M
authors.lastname: Fendrych
authors.affiliation: Department of Plant Systems Biology, VIB, Ghent, 9052, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052, Ghent, Belgium.
authors.forename: A
authors.initials: A
authors.lastname: Goossens
authors.affiliation: Center for the Biology of Disease, VIB, Leuven, Belgium.|Institute for Biology/Genetics, Freie Universität Berlin, Berlin, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Holt
authors.affiliation: Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Department of Respiratory Medicine, Ghent University, Ghent, Belgium.|GROUP-ID Consortium, Ghent University and University Hospital, Ghent, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Janssens
authors.affiliation: Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|VIB Department of Molecular Genetics, Antwerp University, Antwerpen 2020, Belgium.
authors.forename: M
authors.initials: M
authors.lastname: Krols
authors.affiliation: IRIBHM, Université Libre de Bruxelles, Brussels, B-1070, Belgium.
authors.forename: J-C
authors.initials: JC
authors.lastname: Larsimont
authors.affiliation: Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Mc Guire
authors.affiliation: Department of Plant Systems Biology, VIB, Ghent, 9052, Belgium.|Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052, Ghent, Belgium.
authors.forename: M K
authors.initials: MK
authors.lastname: Nowack
authors.affiliation: Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: X
authors.initials: X
authors.lastname: Saelens
authors.affiliation: Carl Zeiss Microscopy, GmbH, Oberkochen, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Schertel
authors.affiliation: Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Schepens
authors.affiliation: Center for the Biology of Disease, VIB, Leuven, Belgium.
authors.forename: M
authors.initials: M
authors.lastname: Slezak
authors.affiliation: VIB Department of Molecular Genetics, Antwerp University, Antwerpen 2020, Belgium.
authors.forename: V
authors.initials: V
authors.lastname: Timmerman
authors.affiliation: Department of Intensive Care, Leiden University Medical Center, Leiden, The Netherlands.|Johnson and Johnson Pharmaceutical Research and Development, Beerse, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Theunis
authors.affiliation: Department of Intensive Care, Leiden University Medical Center, Leiden, The Netherlands.|Johnson and Johnson Pharmaceutical Research and Development, Beerse, Belgium.
authors.forename: R
authors.initials: R
authors.lastname: VAN Brempt
authors.affiliation: Department of Intensive Care, Leiden University Medical Center, Leiden, The Netherlands.|Johnson and Johnson Pharmaceutical Research and Development, Beerse, Belgium.
authors.forename: Y
authors.initials: Y
authors.lastname: Visser
authors.affiliation: VIB Bio Imaging Core, Gent, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Inflammation Research Center, VIB, Technologiepark 927, Gent, B-9052, Belgium.|Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: C J
authors.initials: CJ
authors.lastname: Guérin
mesh_terms: D000818:Animals; D001921:Brain / Q000648:ultrastructure; D055032:Electron Microscope Tomography / Q000379:methods; D016591:Histocytological Preparation Techniques / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008168:Lung / Q000166:cytology / Q000648:ultrastructure; D051379:Mice; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning / Q000295:instrumentation / Q000379:methods*; D008867:Microtomy; D018517:Plant Roots / Q000648:ultrastructure
keywords: Correlative light and electron microscopy; focused ion beam scanning electron microscopy; sample preparation; serial block-face scanning electron microscopy