• Applied Microscopy
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• v.42 no.1
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• pp.49-52
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• 2012

For TEM study of biological samples or polymers that are contained in organic structure, it is often required that the sample is prepared by using ultramicrotome and stained with proper agents to increase the contrast of organic structure. In this study, we investigated an efficient TEM sample preparation method for ductile heterogeneity material by using ultramicrotomy. Cryo-ultramicrotomy is a suitable method that is capable of rendering sample hardness for various ductile materials. However, it has several factors to consider, such as experimental cost, working time and finding the optimal staining conditions. To satisfy these considerations, we prepared TEM sample by using ultramicrotome without cryofunction, and secured the sample hardness by applying the staining process prior to ultrathin sectioning. The cross-linked polyethylene structure in the sample was stained with the 2% $RuO_4$ solution in a sealed test tube for 24 hours at $4^{\circ}C$. After the sample staining, ultrathin sections of sample were prepared using ultramicrotome. As a result, it was revealed that the difficulties associated with staining of ultrathin sections prepared by low-temperature conditions were improved. In addition, appropriate staining depth of sample could be selected for sectioning process. The quality of TEM sample obtained by using this method was better than that of cryo-ultramicroscopy. Finally, it is expected that our method could be effectively applied in TEM sample preparation for a variety of nano-bio convergence materials.

• Applied Microscopy
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• v.48 no.2
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• pp.49-53
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• 2018

A successful transmission electron microscope (TEM) analysis is closely related to the preparation of the TEM specimen and should be followed by the suitable TEM specimen preparation depending on the purpose of analysis and the subject materials. In the case of the Si-based anode material, lithium atoms of formed Li silicide were removed due to ion beam and electron beam during TEM specimen preparation and TEM observation. To overcome the problem, we proposed a new technique to make a TEM specimen without the ion beam damage. In this study, two types of test specimens from the Si-based anode material of Li-ion battery were prepared by respectively adopting the only focused ion beam (FIB) method and the new FIB-ultramicrotome method. TEM analyses of two samples were conducted to compare the Ga ion damage of the test specimen.

• 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.

• The Korean Journal of Zoology
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• v.26 no.4
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• pp.271-282
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• 1983

The authors observed the ultrastructure of the pigment cells of the frog, Rana nigromaculata Hallowell, during the hibernation. The specimens from the skin were fixed in 2.5% glutaraldehyde-paraform-aldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 2% osimium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with LKB-ultramicrotome. the ultrathin sections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follows. In hibernating phase, pigment cells of the frog were consisted of the three kinds of chromatophores (xanthophore, iridophore and melanophore) in their dorsal skin. The traits of these cells were as follows. 1. Xanthophores A. Xanthophores were filled with pterinosomes and carotenoid vesicles. Many ribosomes, a few mitochondria and glycogen particles were dispersed in the cytoplasm. B. Pterinosomes were spherical or ellipsoidal in shape. They were divided into 6 types (type I, type II, type III, type IV, type V, type VI pterinosomes) by the their inner structure and especially, type I, type II, type III pterinosomes were well developed.

• The Korean Journal of Zoology
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• v.22 no.3
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• pp.103-114
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• 1979

The authors observed the ultrastructure of the granular glands in the amphibian skin with an electron microscope. The specimens from the experimental animals (Bombina orientalis, Bufo bufo gargarizans, Rana nigromaculata and Rana rugosa) were fixed in 2.5% glutaraldehyde-paraformaldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 1% osmium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with a LKB-ultramicrotome. the ultrathin sections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follws: 1. The granular gland in the amphibian skin consisted of the glandular epithelial and the myoepithelial cells. 2. The epithelial cells of the granular gland in the amphibian skin consisted of the dark cells but the light cells were also observed in that of Bombina orientalis. 3. The granular glands of the amphibian skin were in holocrine fashion. 4. The nuclei of the epithelial cells of the amphibian cutaneous granular glands were round or oval and showed small and large inforldings of nuclear envelope. Heterochromatins were mainly distributed near the nuclear envelope. Mitochondria were mainly distributed in the perinuclear portion and rough-surfaced endoplasmic reticulums were developed in the cytoplasm but smooth-surfaced endoplasmic reticulums were not well developed. 5. Secretory granules were round or oval and electron-dense and less electron-dense granules were observed. 6. The authors infer that the differences in electron density of the secretory granules in the granular glands of the amphibian skin are due to difference in the concentrations of secretory substances as related to the processes of its formation, and that those chemical components are identical.

• The Korean Journal of Zoology
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• v.21 no.1
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• pp.29-39
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• 1978

The authors observed the ultrastructure of the mucous glandular epithelial cells in the amphibian skin by mean of electron microscope. The specimens from the experimental animals were fixed in 2.5% glutaraldehyde-oaraformaldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 1% osmium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with Sorvall MT-2 ultramicrotome. The ultrasections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follows: 1. The cutaneous mucous glands in amphibia consisted of the glandular epithelial and the myoepithelial cells. 2. Several different cells in ultrastructure were observed in the mucous glandular epithelium of the adult amphibian skin. a. The dark and the light cells were observed in Hynobius leechi. b. The mitochondria-rich and the round secretory granule-containing cells were observed in Bombina orientalis. c. The round secretory granule-containing and the foam-like granule mass-containing cells were observed in Kaloula tornieri. d. The cutaneous mucous gland of Rana nigromaculata were divided into two types: A and B-type glands. In the A-type mucous gland, the mitochondria-rich and the round secretory granule-containing cells and in the B-type mucous gland, the mitochondria-rich, the secretory granule-containing and the ER-rich cells were observed. 3. Based upon the above findings, the authors infer that the mucous granular epithelium of the amphibian skin consists of the mitochondria-rich undifferentiated, the secetory granule-containing and mature, and the ER-rich evacuated cells.

• The Journal of the Korean dental association
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• v.9 no.12
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• pp.819-822
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• 1971

For this investigation, author isolated Streptococcus mitis strain SD-9 from the bacterial flora in the human dental plaque, which was incubated in brain-heart infusion media containing 5% sucrose at 37℃ for 24 hours. For the cytochemical demonstration of polysaccharide produced by this strain, a modified thiosemicarbazide osmium method (Critchley et al., 1967) was used. After fixation with this reagent, the harvested cells was suspended in 1% agar for the higher concentration of cells(Kellenberger et al., 1964). And they were dehydrated in the various concentration of ethanol, and embedded in Epon 812(Luft, 1961). Sectioning was done with the Sorvall MT-2 Porter Blum ultramicrotome by means of a glass knife, and the sections were stained with saturated uranyl acetate and lead citrate (Raynolds, 1963). All preparations were examined in a electron microscope, Hitachi HU-ll E-1 type. The morphological features of extracellular polysaccharide produced by Streptococcus mitis strain SD-9 were appeared in 3 structurally different forms, those are, electron dense fibrillar material linearly arranged adjacent to the outer surface of cell wall, highly electron dense globular material adjacent to the outer surface of cell wall, and strutureless fluffy meshwork of possible very fine filament.

• Parasites, Hosts and Diseases
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• v.22 no.1
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• pp.30-36
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• 1984

An ultrastructural study on the mature spermatoBoa oi Cloptorchis sineitsis was carried out. For this study, the liver cukes were collected from the livers of rabbits and rats artificially infected with the metacercariae obtained from the fresh water fish, Pseudorasbora parve. Six-month old worms were used. The collected liver fiukes were washed with 0.85% saline solution and then immediately moved to cold 2% glutaraldehyde buffered with 0.1M Millonig's phosphate buffier (pH 7.4) . The materials were dissected into appropriate pieces in the fixative about 30 minutes after beginning of the fixation. Two hours later the materials containing the seminal receptacle were rinsed several times with the buffier and were secondarily fixed with cold, bugeyed 1% osmium tetroxide(OsO4) for 2 hours. The fully mixed tissue blocks Ivere dehydrated in a series of graded concentrations of acetone and were embedded in Epon 812 mixture. Thin sections obtained from LKB-5 ultramicrotome were stained with uranyl acetate and Reynold's lead citrate. Observations of the sections were carried out with JEM-100CX II electron microscope, In general, the mature sperm was long thread-like form with a sickle-shaped head. According to the longitudinal sectioned view of the sperm tail, the nucleus seemed to be spirally coiled and run a little far along the tail. The acrosome was not observed. The cytoplasm of the tail was biflagellated as usual in trematodes. Unlike other platyhelminth spermatozoa, the sperm tail of Clenorchis sinensis showed the ${\ulcorner}9+2{\lrcorner}$ in the microtubular arrangement. The mitochondria with poorly developed cristae were observed throughout the middle piece. The middle piece of the tail showed dull ladder or triangular shapes with the two flagella at the bottom. But, the principal piece of the tail was slightly flattened cylindrical shape with two aagella within the cytoplasm. The end piece was uniflagellated. It was not clearly identised whether the end piece was subdivided into two by aagellum or the lengths of the two aagella were different. The glycogen granules were rich in the cytoplasm throughout the lenght of the spermatozoa. These granules might be the energy source for the movement of the spermatosoa.

• Korean Journal of Materials Research
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• v.11 no.4
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• pp.329-334
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• 2001

Microstructures of barrier-type oxide layers on aluminum was studied by XRD, TEM and RBS. Fer formation of oxide layer. aluminum was anodized in a boric acid solution. The thickness of the oxide film subjected to applied voltage increased linearly at ratio of 1.54nm/V. For oxide layer anodized at 300V, amorphous structure of oxide layer was not transformed after heat treatment at 50$0^{\circ}C$ , while for oxide layers anodized at higher voltages the amorphous structure crystallized into a ${\gamma}$-alumina without any heat treatment. It was also found that the amorphous structure of oxide layer formed at 100V transformed into crystalline structure by electron irradiation. The structure was identified as ${\gamma}$-alumina.

• Applied Microscopy
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• v.25 no.1
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• pp.1-14
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• 1995

The purpose of this study was to investigate the main sensory trigeminal nucleus in the aging rat brain by means of electron microscope. Male Sprague-Dawley rats, two (control group) and thirty six (aging group) months of age, were used. These animals were sacrificed by perfusion fixation with 2.5% glutaraldehyde-2.0% paraformaldehyde (0.1M phosphate buffer, pH 7.4) under sodium pentobarbital. The objective area was punched out with a sharp-edged metal cylinder of 0.8 mm in diameter. These blocks of tissue were then washed in 0.1M phosphate buffer, postfixed in 2% osmium tetroxide, dehydrated in a graded series of ethyl alcohol, and embedded in Epon 812. Thin sections were cut with Super Nova ultramicrotome, pick up on grids and double stained with lead citrate and uranyl acetate, and observed in JEOL 100B electron microscope. The results were as follows: 1. In the control group, the neuronal cell body of the main sensory trigeminal nucleus was filled with nucleus, Golgi complex, Nissl substance, mitochondria, microfilaments and microtubules. However, few Nissl substances are seen in neuronal cell body. Axoaxonic synapse, axodendritic synapse, axosomatic synapse, axospinous synapse, myelinated and unmyelinated nerve fibers were well organized around cell bodies. Neurons with abnormal changes were not seen. 2. In the aging group, the neuronal cell body of the main sensory trigeminal nucleus contained large number of lipofuscin granules, dense body and swollen mitochondria. Terminal boutons contained glycogen, crystal-like vesicle and membranous indicating first signs of degeneration. The dendrites were found to be in synaptic contact with altered axon terminals. Frequently axons filled with dark axoplasn and splitted myelin sheath were noticed.