Reconstruction of Neural Circuits Using Serial Block-Face Scanning Electron Microscopy
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초록
Electron microscopy is currently the only available technique with a spatial resolution sufficient to identify fine neuronal processes and synaptic structures in densely packed neuropil. For large-scale volume reconstruction of neuronal connectivity, serial block-face scanning electron microscopy allows us to acquire thousands of serial images in an automated fashion and reconstruct neural circuits faster by reducing the alignment task. Here we introduce the whole reconstruction procedure of synaptic network in the rat hippocampal CA1 area and discuss technical issues to be resolved for improving image quality and segmentation. Compared to the serial section transmission electron microscopy, serial block-face scanning electron microscopy produced much reliable three-dimensional data sets and accelerated reconstruction by reducing the need of alignment and distortion adjustment. This approach will generate invaluable information on organizational features of our connectomes as well as diverse neurological disorders caused by synaptic impairments.
Abstract
Electron microscopy is currently the only available technique with a spatial resolution sufficient to identify fine neuronal processes and synaptic structures in densely packed neuropil. For large-scale volume reconstruction of neuronal connectivity, serial block-face scanning electron microscopy allows us to acquire thousands of serial images in an automated fashion and reconstruct neural circuits faster by reducing the alignment task. Here we introduce the whole reconstruction procedure of synaptic network in the rat hippocampal CA1 area and discuss technical issues to be resolved for improving image quality and segmentation. Compared to the serial section transmission electron microscopy, serial block-face scanning electron microscopy produced much reliable three-dimensional data sets and accelerated reconstruction by reducing the need of alignment and distortion adjustment. This approach will generate invaluable information on organizational features of our connectomes as well as diverse neurological disorders caused by synaptic impairments.
참고문헌 (18)
- Blackstad T W, "Electron microscopy and biological structure research", Tidsskr. Nor. Laegeforen, 85, pp.97~103 (1965)
- Briggman K L, "Volume electron microscopy for neuronal circuit reconstruction", Curr. Opin. Neurobiol, 22, pp.154~161 (2012)
- Albert Cardona, "An Integrated Micro- and Macroarchitectural Analysis of the Drosophila Brain by Computer-Assisted Serial Section Electron Microscopy", PLoS Biology, 8(10), pp.e1000502~ (2010), Public Library of Science (PLoS)[DOI]
- Denk W, "Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure", PLoS Biol, 2, pp.e329~ (2004)
- Frotscher M, "Fine structure of synapses on dendritic spines", Front. Neuroanat, 8, pp.94~ (2014)
- Hayworth K J, "Imaging ATUM ultrathin section libraries with WaferMapper: a multi-scale approach to EM reconstruction of neural circuits", Front. Neural Circuits, 8, pp.68~ (2014)
- Jones C, "Effi cient semi-automatic 3D segmentation for neuron tracing in electron microscopy images", J. Neurosci. Methods, 246, pp.13~21 (2015)
- 김현욱, "Electron Tomography and Synapse Study", 한국현미경학회지, 44(3), pp.83~87 (2014), 한국현미경학회
- Knott G, "Serial section scanning electron microscopy of adult brain tissue using focused ion beam milling", J. Neurosci, 28, pp.2959~2964 (2008)
- 이계주, "고압전자현미경을 이용한 소뇌 평행섬유-조롱박세포 간 신경연접의 3차원 재구성", 한국현미경학회지, 35(1), pp.31~39 (2005), 한국현미경학회
- Lee K J, "Efficient three-dimensional reconstruction of synapse with high-voltage electron microscopy", J. Electron Microsc, 54, pp.139~141 (2005)
- Lee K J, "Motor skill training induces coordinated strengthening and weakening between neighboring synapses", J. Neurosci, 33, pp.9794~9799 (2013)
- Marx V, "Neurobiology : brain mapping in high resolution", Nature, 503, pp.147~152 (2013)
- Mishchenko Y, "Reconstruction of complete connectivity matrix for connectomics by sampling neural connectivity with fluorescent synaptic markers", J. Neurosci. Methods, 196, pp.289~302 (2011)
- Nguyen H B, "Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging", Sci. Rep, 6, pp.23721~ (2016)
- Steward O, "Preferential localization of polyribosomes under the base of dendritic spines in granule cells of the dentate gyrus", J. Neurosci, 2, pp.284~291 (1982)
- Walton J, "Lead aspartate, an en bloc contrast stain particularly useful for ultrastructural enzymology", J. Histochem. Cytochem, 27, pp.1337~1342 (1979)
- Wilke S A, "Deconstructing complexity: serial block-face electron microscopic analysis of the hippocampal mossy fi ber synapse", J. Neurosci, 33, pp.507~522 (2013)