1.Localization the antigens in cells by immunoelectronmicroscopic method
Journal of Practical Medicine 2004;472(2):14-17
In the study, indirect immunogold labelling technique was used after embedding on verocells infected by measle virus and on PMK cells infected by rotavirus. According to black particles in immuno-electronic microscopic figure, rotavirus in PMK cells was localized and the formation of measle virus in vero cell was realized.
34.
Trần Thanh Dương, Nguyễn Thu Vân, Hoàng Thủy Long: Molecula epidemiology of C hepatitis in hepatitis patients in Hanoi city. Vn J Pract Med 2004; 472(2):17-22:(CIMSI)
Epidemiology; Hepatitis C; Fibrosis; Epidemiology, Molecular
323 patients with hepatitis and cirrhosis (244 males, 89 females admitted into the clinical Institute of Tropical diseases from Jan 2001 to August 2002 were studied on. Result showed that: in Hanoi, hepatitis patients were infected with C hepatitis virus of the types 1a, 1b, 6a. The incidence of various HCV genes was not different in deverse age groups, places, occupations, marital status of the patients, but the differences were noted according to the gender, the education level, the history of drug use and blood transfusion.
Antigens
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Cells
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Microscopy, Immunoelectron
2.Immunoelectronmicroscopic localization of antigen in cells
Journal of Medical Research 2003;26(6):6-11
Immunoelectronmicroscopic examination was applied largely in the diagnosis of microbioorganism. The technique of indirect immunogolden labelling was used after embedding on vero cell infected with measle virus and on TVP cell infected with rotavirus SA11. By black particles in immunoelectromicroscopic figure, the localization of rotavirus in TVP cell was determinated and the formation of measle virus in vero cell was identified.
Antigens
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cells
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Microscopy, Immunoelectron
3.Immunoelectron-microscopic localization of IgE binding site of mugwort pollen.
Hae Sim PARK ; Jae Won KIM ; Chein Soo HONG
Journal of Korean Medical Science 1993;8(1):30-33
To elucidate the IgE binding site of mugwort (Artemisia vulgaris r.) pollen, pollen grains were frozen and fixed using a cryocut. They were incubated with antibodies according to the following sequence: Sera pool of individuals who showed mugwort-RAST class 3 or 4, biotin-labeled goat anti-human IgE antibody, streptavidin-peroxidase and diaminobenzidine. Then, they were observed under electron microscopy. The control section was incubated with the sera pool from individuals who showed a negative result on a skin prick test to mugwort pollen. Antigenic activity (electrondense line) was noted on the surface of the exine. There was no activity in cytoplasm or the intine layer. The control section was completely free of activity. It was suggested that the IgE binding site of mugwort pollen was present on the surface of the exine.
Binding Sites
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Humans
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Immunoglobulin E/*metabolism
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Microscopy, Electron
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Microscopy, Immunoelectron
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Pollen/*immunology
5.Detection of Helicobacter pylori by Pre-embedding Immunoelectron Microscopy: Comparison with Immunoblotting Method.
Hae Seoung JUNG ; Eun Jung KIM ; Eun A KIM ; Ji Hoe PARK ; Jin Su JUN ; Ji Hyoun SEO ; Jae Young LIM ; Myoung Bum CHOI ; Hyang Ok WOO ; Hee Shang YOUN ; Gyung Hyuck KO ; Seung Chul BAIK ; Woo Kon LEE ; Myung Je CHO ; Kwang Ho RHEE
Journal of the Korean Pediatric Society 2002;45(7):862-874
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 and E) staining, and one for pre- embedding 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 and 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 and 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.
Biopsy
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Child
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Diagnosis
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Helicobacter pylori*
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Helicobacter*
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Humans
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Immunoblotting*
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Microscopy, Immunoelectron*
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Urease
6.An immunohistochemical study on the initial development of 155 kd neurofascin in rat sciatic nerve.
Hee Young WON ; Ik Hyun CHO ; Jong Hwan LEE ; Sang Soep NAHM ; Nong Hoon CHOE ; Jae Hyung PARK ; Byung Joon CHANG
Korean Journal of Veterinary Research 2012;52(3):199-203
Neurofascin, one of the members of L1CAM, has been known to have some important roles during the development of nerve fibers. In order to investigate the role of neurofascin associated with the development of nerve fibers in the rat sciatic nerve, the initial development of NF155 in the paranode was studied with immuno-fluorescence and immuno-electron microscopy. The result of the present study showed NF155 was not detected in the fetal sciatic nerve and began to reveal at the postnatal day 0 (P0) and dramatically increased by time lapse until postnatal day 7 (P7). NF155 was prominently localized in the axolemma of paranode and not detected in the central region of node of Ranvier. According to the present study, NF155 is likely to have some relationships with the formation of paranode and myelin sheath.
Animals
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Immunohistochemistry
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Microscopy, Immunoelectron
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Myelin Sheath
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Nerve Fibers
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Neural Cell Adhesion Molecule L1
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Rats
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Sciatic Nerve
7.A Pre-embedding Technique of Nanogold-Silver Staining for Immunocytochemical Localization of Gp100 in Cultured Melanocytes.
Korean Journal of Anatomy 2002;35(1):75-82
Nanogold-silver staining is a pre-embedding immunocytochemical technique, making it possible to label antigens with particulate markers at the electron microscopic level. This technique is based on the advancement of gold technology and on the development of silver staining or enhancement. In the present study, the method of Nanogoldsilver staining that includes the use of high concentration of glutaraldehyde and gold toning as well, was used to localize gp100 or a membrane bound antigen in melanosome of the cultured melanocytes. With the HMB45 antibody against gp100, biotinylated secondary antibody and streptavidin-nanogold followed by silver enhancement and gold toning led to highly specific labeling of gold particle over the melanosome compartments. The specificity of labelings obtained with this protocol was confirmed by the control experiment. Omission of the primary antibody led to very low background of labeling. The specific signal that appeared as a collection of gold particles was localized mainly to the peripheral part of melanosome. This finding provides the more detailed nature of gp100 localization in melanosome, which has not yet been shown by the previous immunocytochemical study such as the post-embedding immunoelectron microscopy.
Glutaral
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Melanocytes*
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Melanosomes
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Membranes
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Microscopy, Immunoelectron
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Sensitivity and Specificity
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Silver
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Silver Staining
8.Immunoelectron microscopic localization of partially purified antigens in adult Paragonimus iloktsuenesis.
The Korean Journal of Parasitology 2001;39(2):119-132
An immunoelectron microscopy employing immunogold labeling method was performed to detect tissue origin of D1 fraction (D1A) 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. D1A 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 D1A was mainly originated from the intestinal epithelial tissues before the development of vitelline gland of the parasites. Immuno-reactivity of two immunoglobulins (PI-Ig, D1-Ig) was significantly different in intestinal epithelial cytoplasmic protrusions (CP) and intestinal epithelial secretory granules (SG). In the experimental group with D1-Ig, gold particles were labeled significantly in CP than in SG when compared to the PI-Ig group. Thus, the major antigenic materials in D1 antigen having a strong antigenicity in the early infection period was considered to be originated from the intestinal epithelial tissue.
Animals
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Antigens, Helminth/*analysis/isolation & purification
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Chromatography, Ion Exchange
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Immunohistochemistry/methods
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Microscopy, Immunoelectron
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Paragonimus/*immunology
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Support, Non-U.S. Gov't
9.Expression and significance of alpha5beta1 integrin in abnormal scars.
You YU ; Guang-Zhao HE ; Yi WU ; Chong-Ben HUANG
Chinese Journal of Plastic Surgery 2004;20(4):271-273
OBJECTIVETo study the expression of alpha5beta1 integrin in the abnormal scars and its role and significance in the formation and development of abnormal scars.
METHODSThe expression of alpha5beta1 integrin was observed in hypertrophic scar (15 samples), keloid (15 samples) and normal skin (10 samples) with SP immunohistochemical method and colloidal gold immuno-electron microscopic technique. The data were semi-quantitatively analyzed.
RESULTSThe expression levels of alpha5beta1 integrin in the fibroblasts of keloids and hypertrophic scars were higher than normal skin; the expression of alpha5beta1 integrin in the fibroblasts of keloids was higher than hypertrophic scars (P < 0.01).
CONCLUSIONThe alpha5beta1 integrin appears to have close relation to the formation and development of abnormal scars. To find a way to decrease the expression level of alpha5beta1 integrin in fibroblasts may be a new approach to inhibit scar hypertrophy.
Cicatrix ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Integrin alpha5beta1 ; analysis ; metabolism ; Keloid ; metabolism ; pathology ; Microscopy, Immunoelectron ; Skin ; chemistry ; pathology ; ultrastructure
10.Ultrastructural localization of 28 kDa glutathione S-transferase in adult Clonorchis sinensis.
Sung Jong HONG ; Jae Ran YU ; Shin Yong KANG
The Korean Journal of Parasitology 2002;40(4):173-176
Glutathione S-transferase (28GST) with molecular mass of 28 kDa is an antioxidant 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 bioreactive 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.
Animals
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Clonorchis sinensis/*enzymology
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Glutathione Transferase/*metabolism/physiology
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Immunohistochemistry
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Microscopy, Immunoelectron
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Molecular Weight
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Support, Non-U.S. Gov't