• Applied Microscopy
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• v.48 no.4
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• pp.87-95
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• 2018

Immunoelectron microscopy using an antigen-antibody reaction in an electron microscope is a very useful tool to identify the components of a tissue in an electron microscope. Many researchers also use immunoelectron microscopy. Nonetheless, immunoelectron microscopy is rarely introduced systematically, and immunoelectron microscopy can be carried out without fully understanding the principles, and cases of poor understanding can often be seen in the vicinity. Therefore, in order to make it easier to understand, we will first introduce the principles of immunoelectron microscopy and describe practical methods.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.575-581
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• 1995

Both immunoelectron microscopy(IEM) and immunogold conjugate immunoelectron microscopy (IGC-IEM) techniques were developed for the detection of porcine epidemic diarrhea virus(PEDV) from the feces. Fecal samples were incubated sequentially with anti-PEDV monoclonal antibody(MoAb) and immunogold conjugated goat anti-mouse IgG+IgM. Then negatively stained, mounted on the formvar carbon-coated copper EM grids and observed by the transmission electron microscope. By the direct electron microscopy(DEM), coronavirus particles were observed from 17 cases of total 33 fecal samples of grower pigs and sows. The virons of coronavirus were moderately pleomorphic but mostly spherical, with a diameter ranged from 90 to 190nm. PED virus particles were identified from 15 cases of 17 DEM positive samples by the IEM and IGC-IEM techniques. Aggregates of PED virus coated with specific antibody were seen in fecal samples incubated with homologous anti-PED virus MoAb but not in control samples incubated with anti-TGE virus MoAb. Following incubation with immunogold-conjugated secondary antibody, the gold granules were usually distributed around and among the virus particles and soluble and viral particle-associated antigen. So, IEM and IGC-IEM techniques were proved a rapid and sensitive methods for detection and identification of PED virus from fecal and intestinal contents.

• Clinical and Experimental Pediatrics
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• v.45 no.7
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• pp.862-874
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• 2002

Purpose : We tried to evaluate whether the detection rate of Helicobacter pylori in gastric biopsy specimens could be improved by using pre-embedding immunoelectron microscopy. Methods : A total of 119 children who complained of upper gastrointestinal symptoms were endoscoped at the Gyeongsang National University Hospital from July, 1996 to July, 1999. Five biopsy specimens(three for urease test, one for hematoxylin-eosin(H & E) staining, and one for preembedding immunoelectron microscopy) were obtained from each antrum and body. Immunoblotting analysis were also performed. Results : Among the 119 patients, H. pylori were found in 116 patients(97.5%) by the immunoelectron microscopy. Among three patients who were found H. pylori negative in immunoelectron microscopy, two patients showed H. pylori in H & E stained slides and one patient was urease test positive(color change within six hours). Urease tests were positive in 107 patients(89.9 %). The positive rate of immunoblotting tests was 81.5%. However, only 13 patients(10.9%) showed H. pylori on the H & E stained antrum or body tissue. Conclusion : In this study, we found H. pylori histopathologically in most of the pediatric patients who complained of upper gastrointestinal symptoms. This study showed that pre-embedding immunoelectron microscopic examinations can be used as a gold standard in the diagnosis of childhood H. pylori infection. However, this method also has limited capacity to detect widely scattered H. pylori compared to the other histopathologic diagnostic methods.

• Parasites, Hosts and Diseases
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• v.39 no.2
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• pp.119-132
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• 2001

An immunoelectron microscopy employing immunogold labeling method was performed to detect tissue origin of Dl fraction (DIA) among 5 antigenic protein fractions partially purified by DEAE- anion exchange chromatography from water- soluble crude antigen (PIWA) of adult Paragonimus iloktsuenensis. Immune reactions of adult worm tissues with rabbit serum immunoglobulin immunized with crude antigen (PI-Ig) and D1 antigen (D1-Ig), as well as rat serum immunoglobulin infected with P. iloktsuenensis were observed. DlA showed strong antigenicity in the intestinal epithelium of the worms during the early infection period of 2-4 weeks after infection. The vitellaria also showed stronger antigenicity than the other tissue sites in immune reaction of tissues against all immunoglobulins from 4 to 33 weeks after vitelline development. Therefore, it is suggested that DlA was mainly originated from the intestinal epithelial tissues before the development of vitelline gland of the parasites. Immune-reactivity of two immunoglobulins (PI-Ig, Dl-Ig) was significantly different in intestinal epithelial cytoplasmic protrusions (CP) and intestinal epithelial secretory granules (SG). In the experimental group with Dl-Ig, gold particles were labeled significantly in CP than in SG when compared to the PI-Ig group. Thus, the major antigenic materials in Dl antigen having a strong antigenicity in the early infection period was considered to be originated from the intestinal epithelial tissue .

• Korean Journal of Microbiology
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• v.40 no.1
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• pp.23-28
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• 2004

Antigenic analysis of Herpes simplex type 2 virus was performed and its major antigen was localized using an immunoelectron microscopy. Antigens of 32, 43, 59 and 69 kDa were constantly expressed during the course of infection for 48 hr in the infected Vero cell. An antigen of 51 kDa was turned out to be the major one in inducing a immune response in Western-blot analysis. The 51 kDa antigen was localized on the surface of HSV-2 by immunoelectron microscopy using colloidal golds and anti-HSV 2 polyc1onal antibody. Immunofluorescence assay indicated that viral antigens were found throughout the infected cell and, especially, on the surface of the cell.

• Applied Microscopy
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• v.28 no.4
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• pp.465-475
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• 1998

The cryoprotection, section retrieval and embedding methods were studied for the preservation of ultrastructure of ultracryomicrosections in immunoelectron microscopy. The results obtained were as follows. 1. The cryoprotection of ultrastructure with a mixture containing 1.7 M sucrose and 15% polyvinylpyrrolidone was better than that with 2.3 M sucrose. The stretching caused by surface tension and the electron lucent holes decreased more in the cryosections infused with 2.3 M sucrose than in those with the mixture. 2. The difference between section retrieval solutions in cases of cryoprotection with 2.3 M sucrose was that the destructive .effects such as electron lucent holes and stretching between myofribrils were less in a mixture containing 1% methylcellulose and 2.3 M sucrose than in 2.3 M sucrose. The difference was obscure in the mixture containing 1.7 M sucrose and 15% PVP, but the destructive effects were slightly less in a mixture containing 1% mthylcellulose and 2.3 M sucrose than in 2.3 M sucrose or 1% methylcellulose. 3. The embedding of cryosection on drying with 2% PVA or 2% methylcellulose exhibited some protective effect during observation with transmission electron microscope, but made the ultrastructure more obscure. 4. Mitochondrial membrane and cristae and myofilaments were well delinated in sections infused with 2.3 M sucrose and retrieved with 1% methylcellulose and 2.3 M sucrose. In summary, it is suggested that the cryoprotection with 2.3 M sucrose and section retrieval with a mixture containing 1% methylcellulose and 2.3 M sucrose are good for the ultrastructure of cryosections.

• Parasites, Hosts and Diseases
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• v.40 no.4
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• pp.173-176
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• 2002

Glutathione S-transferase (28GST) with molecular mass of 28 kDa is an anti-oxidant enzyme abundant in Clonorchis sinensis. In adult C. sinensis, 28GST was localized in tegumental syncytium, cytons, parenchyma, and sperm tails examined by immunoelectron microscopy. C. sinensis 28GST was earlier found to neutralize bio-reactive compounds and to be rich in eggs. Accordingly. it is suggested that 28GST plays important roles in phase II defense system and physiological roles in worm fecundity of C. sinensis.

• Applied Microscopy
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• v.31 no.4
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• pp.347-354
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• 2001

Immunoelectron microscopy of storage protein at early stage of seed development showed legumin was firstly accumulated protein in between endoplasmic reticulum (ER) cisternae, and these accumulates were differentiated into protein body (PB) by transformation at later stage. Thin sections of pea cotyledons during the later stages of seed maturation showed three morphologically different types of protein bodies. One of these, presented as rough-surfaced cisternae with terminal dilations, which contained protein deposits and were often found interdigitated between stacks of rough endoplasmic reticulum. Conventional electron microscopy at earlier stages of cotyledon development showed this protein body type initially developed from the rough ER. This transformation of endoplasmic reticulum into a protein body is believed to represent a new pathway of protein body development.

• Applied Microscopy
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• v.25 no.1
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• pp.122-129
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• 1995

Pancreatic endocrine cells of the native Korean goat were investigated immunocytochemically at electron microscopic level. All glucagon-, insulin-, somatostatin- and pancreatic polypeptide(PP)-immunoreactive cells were showed chromogranin(CG) immunoreactivity in the secretory granules of each cells. In addition, bovine pancreatic polypeptide immunoreactivity was found to be colocalized in the secretory granules of the glucagon and insulin cells. These observations support that chromogranin is available as the marker of pancreatic endocrine cells on the native Korean goat and BPP colocalized in the secretory granules of glucagon and insulin cells.

• Korean Journal of Veterinary Research
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• v.38 no.3
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• pp.488-496
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• 1998

Somatotropes, mammotropes and somatomammotropes of the Korean native goat hypophysis were studied by double immunoelectron microscopy using antisera to growth hormone(GH) and prolactin(PRL), and protein A-gold particles of different sizes. Mammotropes were round or oval in shape, and contained round and electron dense secretory granules. The size of secretory granules was variable from 460nm to 680nm in diameter. Somatotropes were elliptical or triangular in shape and the oval nucei were located eccentrically at the periphery of the cell. Secretory granules of the cell were oval in shape and clearly distinguished from round granules of mammotropes. The size of granules was 320~680nm in diameter, smaller than that of mammotropes. Somatomammotropes contained round or oval secretory granules. The granules had intermediate size between somatotropes and mammotropes. Some of granules contained both GH and PRL, while the others contained only one of them.