title: Challenges of microtome-based serial block-face scanning electron microscopy in neuroscience.
abstract: Serial block-face scanning electron microscopy (SBEM) is becoming increasingly popular for a wide range of applications in many disciplines from biology to material sciences. This review focuses on applications for circuit reconstruction in neuroscience, which is one of the major driving forces advancing SBEM. Neuronal circuit reconstruction poses exceptional challenges to volume EM in terms of resolution, field of view, acquisition time and sample preparation. Mapping the connections between neurons in the brain is crucial for understanding information flow and information processing in the brain. However, information on the connectivity between hundreds or even thousands of neurons densely packed in neuronal microcircuits is still largely missing. Volume EM techniques such as serial section TEM, automated tape-collecting ultramicrotome, focused ion-beam scanning electron microscopy and SBEM (microtome serial block-face scanning electron microscopy) are the techniques that provide sufficient resolution to resolve ultrastructural details such as synapses and provides sufficient field of view for dense reconstruction of neuronal circuits. While volume EM techniques are advancing, they are generating large data sets on the terabyte scale that require new image processing workflows and analysis tools. In this review, we present the recent advances in SBEM for circuit reconstruction in neuroscience and an overview of existing image processing and analysis pipelines.
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, 4058, Basel, Switzerland.
authors.forename: A A
authors.initials: AA
authors.lastname: Wanner
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, 4058, Basel, Switzerland.
authors.forename: M A
authors.initials: MA
authors.lastname: Kirschmann
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, 4058, Basel, Switzerland.
authors.forename: C
authors.initials: C
authors.lastname: Genoud
mesh_terms: D000818:Animals; D001921:Brain / Q000648:ultrastructure; D063132:Connectome; D016591:Histocytological Preparation Techniques; D021621:Imaging, Three-Dimensional / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000295:instrumentation / Q000379:methods*; D008867:Microtomy; D009434:Neural Pathways / Q000648:ultrastructure*; D009474:Neurons / Q000648:ultrastructure; D009488:Neurosciences / Q000379:methods*; D013569:Synapses / Q000648:ultrastructure
keywords: Microtome; SBFSEM; registration; segmentation; serial block-face scanning electron microscopy (SBEM); three-dimensional reconstruction; tiling