• Title/Summary/Keyword: Correlative light and volume electron microscopy

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Nano-Resolution Connectomics Using Large-Volume Electron Microscopy

  • Kim, Gyu Hyun;Gim, Ja Won;Lee, Kea Joo
    • Applied Microscopy
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    • v.46 no.4
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    • pp.171-175
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    • 2016
  • A distinctive neuronal network in the brain is believed to make us unique individuals. Electron microscopy is a valuable tool for examining ultrastructural characteristics of neurons, synapses, and subcellular organelles. A recent technological breakthrough in volume electron microscopy allows large-scale circuit reconstruction of the nervous system with unprecedented detail. Serial-section electron microscopy-previously the domain of specialists-became automated with the advent of innovative systems such as the focused ion beam and serial block-face scanning electron microscopes and the automated tape-collecting ultramicrotome. Further advances in microscopic design and instrumentation are also available, which allow the reconstruction of unprecedentedly large volumes of brain tissue at high speed. The recent introduction of correlative light and electron microscopy will help to identify specific neural circuits associated with behavioral characteristics and revolutionize our understanding of how the brain works.

Current Status of Automatic Serial Sections for 3D Electron Microscopy

  • Choi, Hyosun;Jung, Min Kyo;Mun, Ji Young
    • Applied Microscopy
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    • v.47 no.1
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    • pp.3-7
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    • 2017
  • The automatic equipment for three-dimensional electron microscopy (3DEM) can acquire serial sections of a large sample in a relatively short time, and is especially suitable for the connectomics, which is a field related to understanding the brain structure as a whole. As many results obtained through 3DEM using automatic serial sections have been published in the field of brain research, many researchers continue to apply this technique to various samples. We reviewed the equipment for automatic serial sectioning, the block preparation method, the limitations of 3DEM, and future directions.

Electron Microscopic Studies on Olfactory Bulbs in the Vertebrates by Phylogenetics (계통발생에 따른 척추동물의 뇌후구에 대한 전자현미경적 연구)

  • Choi, W.B.;Chung, Y.H.;Seo, J.E.
    • Applied Microscopy
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    • v.15 no.2
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    • pp.31-68
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    • 1985
  • Authors are trying to unveil the ultrastructural organization of olfactory bulb, which has been summerized under light microscopic level or communicated only in some detail in different view point until now. For the critical point of view, since the phylogenetical approach will give the ultimate value in the correlative study between structural and functional bases (Brodal, 1969), the present study was carried out light and electron microscopic analyses of the structures of the neurons and synaptic organizations in olfactory bulbs from different animals in phylogenetical scale. We selected each one species from five animal classes: the house rabbit(Oryctolagus cuniculus var. domesticus [Gmelin]) from Mammalia, the domestic fowl (Gallus gallus domesticus Brisson) from Aves, the viper (Agkistrodon hylys [G.P. Pallas]) from Reptilia, a frog (Bombiana orientalis Boulenger) from Amphibia and the crussian carp (Carassius carassius [Linne]) from Pisces. For light microscopic study, samples were fixed in 10% formalin and paraffin sections were stained with hematoxylin-eosin. For the electron microscopic study, the tissues were fixed by perfusion through the heart or immersion with 1% paraform-aldehyde-glutaraldehyde mixture (phosphate buffer, pH 7.4), and final tissue block trimmed under dissecting microscope were osmicated (1% OsO4), they were embedded in Araldite or Epon 812, and ultrathin sections were made by LKB-V ultratome following the inspection of semi-thin sections stained with toluidine blue-borax solution. Ultra-thin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100CX electron microscope. We have summerized our morphological analyses as follows: 1. The olfactory bulb of rabbit, viper and frog shows the eight layers of fila olfactoria, glomerular, external granular, external plexiform, mitral cell, internal plexiform, internal granular, medullary but domestic fowl shows the five layers of glomerular, fibrillar, mitral, granular and medullary and the three layers of fibrilla, glomerular and medullary in crussian carp. The sharpness of demarcation between the layers shows deferential tendency according to phylogenetical order. 2. Mitral cells of vertebrate have large triangular or oval shape with spherical nuclei which contain not so much chromatin. The cytoplasm contains numerous cell organelles, of which Nissl's bodies or granular endoplasmic reticula arranged as parallel strands. Development of granular endoplasmic reticula were declined as the phylogentical grade is going lower. 3. Tufted cells of all animal are mostly spindle or polygonal contour and contain oval nuclei which located in periphery of cytoplasm. The nuclei of rabbit, fowl, viper and frog has relatively space chromatin, but a nucleus of crussian carp contain irregularly aggregated chromatin in karyoplasm. Their cytoplasmic volume and cell organelle contents are in between those of mitral cell and granular cell. They contain moderate amount of mitochondria, granular endoplasmic reticula, a few Golgi complex, polysomes, lysosome, etc. 4. Granule of cells of all the vertebrate amimals studied exhibit similar features; cells and their dense nuclei show spherical or oval contour, and they have the thin rim of cytoplasm which contain only a few cell organelles. 5. In rabbit, the soma of mitral cells were in contact with boutons with two types of synaptic vesicles, that is, round and flat vesicles, especially flat vesicles in boutons were showing reciprocal synapses. However, in domestic fowls, vipers, frogs and crussian carps, there were found boutons showing only spherical synaptic vesicles. 6. The boutons containing round synaptic vesicles were made contact with the some of tufted cell of olfactory bulb in the rabbits, fowls, vipers and frogs, but no synaptic boutons were observed in soma of tufted cells in crussian carps. In the frogs, there were observed dendrites were contact with the soma of tufted cells. 7. In the neuropils of plexiform, granular and glomerular layers olfactory bulbs in the vertebrate, the synapses were axo-large dendrites, axo-median and small dendrites, dendrodendritic, and axo-axonal contacts. However, in the neuropil of crussian carps, synapses were observed only in glomerular layer.

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