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 ; Cells ; Microscopy, Immunoelectron

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 ; cells ; Microscopy, Immunoelectron

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 ; Humans ; Immunoglobulin E/*metabolism ; Microscopy, Electron ; Microscopy, Immunoelectron ; Pollen/*immunology

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 ; Immunohistochemistry ; Microscopy, Immunoelectron ; Myelin Sheath ; Nerve Fibers ; Neural Cell Adhesion Molecule L1 ; Rats ; Sciatic Nerve

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 ; Child ; Diagnosis ; Helicobacter pylori* ; Helicobacter* ; Humans ; Immunoblotting* ; Microscopy, Immunoelectron* ; Urease

Animals ; Hepatitis B virus ; isolation & purification ; Mice ; Mice, Transgenic ; Microscopy, Immunoelectron ; Serum ; virology

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 ; Melanocytes* ; Melanosomes ; Membranes ; Microscopy, Immunoelectron ; Sensitivity and Specificity ; Silver ; Silver Staining

Animals ; Glutamic Acid ; analysis ; toxicity ; Guinea Pigs ; Microscopy, Immunoelectron ; Vestibule, Labyrinth ; chemistry

Podocyte foot processes are interdigitated to form the slit diaphragm and are crucial for the glomerular filtration barrier. Glucocorticoid-induced transcript 1 (GLCCI1) is transcriptionally regulated, but its signaling pathway in podocytes is unknown. The main objective of this study was to investigate the regulation of podocyte foot process proteins and to investigate the role of GLCCI1 in the phosphoinositide 3-kinase (PI3K) pathway using high glucose-induced podocytes and streptozotocin-induced diabetic rats. In podocytes and rat kidneys, GLCCI1 was found to be highly specific for the glomerulus and podocyte foot processes similar to other podocyte-specific proteins (nephrin, podocin, synatopodin and podocalyxin) based on reverse transcription-PCR, western blotting, immunofluorescence and immunoelectron microscopy analyses. In addition, the decrease in the GLCCI1 expression level under hyperglycemic conditions was restored by treatment with a PI3K inhibitor (wortmannin). Immunofluorescence analysis confirmed that GLCCI1 colocalized with nephrin and synaptopodin both in vivo and in vitro. Finally, immunoelectron microscopy data from streptozotocin-induced diabetic rats showed that GLCCI1 also localized in podocyte foot processes. Hence, GLCCI1 is a component of podocyte foot processes, and its expression appears to be regulated via the PI3K pathway.
Animals ; Blotting, Western ; Diaphragm ; Fluorescent Antibody Technique ; Foot* ; Glomerular Filtration Barrier ; In Vitro Techniques ; Kidney ; Microscopy, Immunoelectron ; Podocytes* ; Rats

We examined the fibronectin (FN) secretion of cultured trabecular meshwork (TM) cells in a normal human eye by indirect immunofluorescent technique using mouse anti-human FN monoclonal antibody and FITC-conjugated goat anti-mouse IgG. To localize FN on frozen sections of normal TM, which were obtained from 7 enucleated eyes owing to traumatic eyeball rupture, the same indirect immunofluorescent method was used. Immunoelectron microscopy was applied to demonstrate the distribution pattern of FN in the normal TM of 2 human eyes using an avidin-biotin-peroxidase complex method. In the tissue culture of TM, the TM cell walls and extracellular matrices showed an intense staining with antibody to FN. Indirect immunofluorescent staining of FN on frozen sections of TM showed strong positive reactions in the subendothelial region. There was no reaction in the central core of the trabecular beam. Immunoelectron microscopy revealed the reaction products to FN in the areas lining the trabecular endothelial cells.
Adolescent ; Adult ; Antibodies, Monoclonal ; Cells, Cultured ; Fibronectins/biosynthesis/*metabolism ; Fluorescent Antibody Technique ; Humans ; Male ; Microscopy, Immunoelectron ; Trabecular Meshwork/*metabolism/ultrastructure