• Title/Summary/Keyword: ABC immunohistochemistry

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Immunohistochemistry of Paraffin-embedded Tissues by Super-signal Induction Method (슈퍼시그날 증폭 기술에 의한 파라핀 매몰조직의 면역조직화학염색)

  • Yun Young Gab;Lee Jang Cheon;Jang Seon Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.4
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    • pp.1154-1158
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    • 2004
  • The classical ABC (avidin-biotin peroxidase complex) method for immunohistochemistry in the paraffin-embedded tissues bring into being disadvantage such as low sensitivity of antigen detection and highly background. The biotinyl-tyramide conjugation recently introduced for sensitive immunohistochemistry was applied to light microscopy in paraffin-embedded pancreatic and liver tissues. The protocol consists of an indirect method in which 4-5㎛ tissue sections are reacted successively within a specific primary antibody, followed by a biotinylated secondary antibody, streptavidin-horseradich peroxidase (HRP), and then finally with biotinyl-tyramide. The labeling obtained for insulin and collagen antigen tested in pancreatic and liver tissues, respectively, was found to be highly specific with the labeling for each antigen confined to its particular cellular compartment. In this study, fish (flounder) serum was specially applied to remove nonspecific binding. Background levels and nospecific deposition of the staining were negligible. This results suggest that super-signal induction method by biotinyl-tyramide conjugate can readily applied to antigen detection of the paraffin-embedded tissues.

Congenital transmission of Theileria sergenti in cattle verified by immunohistochemistry (소 Theileria sergenti의 태반감염에 대한 면역세포화학적 증명)

  • Baek, Byeong-kirl;Kim, Jin-ho;Onuma, Misao
    • Korean Journal of Veterinary Research
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    • v.37 no.4
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    • pp.825-829
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    • 1997
  • The spleen and placenta from the aborted fetuses as well as lymphnodes and placenta of the corresponding dams naturally infected with T sergenti were used to localize the parasite antigens by immunohistochemical staining for the possible congenital transmission of theileriosis. Parasite-specific antigens were detected immunohistochemically by incubating the sections with specific monoclonal antibody prepared against 34KD surface antigen of T sergenti and visualized via the avidin biotin complex(ABC) method. Specific T sergenti antigen was detected in the sections of formalin or acetone-fixed fetal spleens and placenta. Similar antigens were also demonstrated in lymphnodes and placentas of the corresponding dams. It is concluded that this technique will eventually play an important role in specialized diagnostic laboratories in the verification/evaluation of congenital infection with T sergenti.

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Association between ABCB1 Immunohistochemical Expression and Overall Survival in Gastric Cancer Patients

  • de Oliveira, Juliana;Felipe, Aledson Vitor;Neto, Ricardo Artigiani;Oshima, Celina Tizuko;de Souza Silva, Marcelo;Forones, Nora Manoukian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6935-6938
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    • 2014
  • Gastric cancer (GC) is one of the most common malignancies worldwide. The ABCB1 protein, a member of the ATP-binding cassette (ABC) transporter family, encoded by the ABCB1 gene, considerably influences the distribution of drugs across cell membranes as well as multidrug resistance (MDR) of antineoplastic drugs. In contrast to the extensive knowledge on the pharmacological action of ABCB1 protein, the correlation between the clinical-pathological data and ABCB1 protein expression in patients with GC remains unclear. The aim was to investigate association between ABCB1 expression and overall survival in GC patients. Human tumor fragments from 57 GC patients were examined by immunohistochemistry assay. We observed lower survival rate of patients with GC who were positive for ABCB1 expression (p=0.030). Based on these observations, we conclude that GC patients with positive ABCB1 protein immunohistochemical expression in their tumors suffer shorter overall survival.

Studies on the rabbit viral hepatitis : Immunohistochemical observations (토끼의 바이러스성 간염에 관한 연구 : 면역조직화학적 관찰)

  • Lee, Cha-soo;Shin, Tae-kyun;Choi, Youn-ju;Jeong, Kyu-sik;Jyeong, Jong-sik
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.269-275
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    • 1993
  • Tissue distribution of RHDV in rabbits were examined by immunofluorescence and ABC methods. Tissues including liver, spleen, kidneys, lungs and brain were frozen, cut in a crycut, and fixed in 10% buffered formalin, embedded in paraplast, and cut $5{\sim}7{\mu}m$ thickness. Sections were immunostained Tissue distribution of RHDV in rabbits were examined by immunofluorescence and ABC methods. Tissues including liver, spleen, kidneys, lungs and brain were frozen, cut in a crycut, and fixed in 10% buffered formalin, embedded in paraplast, and cut $5{\sim}7{\mu}m$ thickness. Sections were immunostained with primary antiserum and conjugated second antibodies as recommended by manufacturer. None of the cultures tested showed virus-induced phenomena. Immunoreactive products were commonly found in the liver, in some cases there were also positive staining in the spleen and kidneys. Other organs showed weak or insignificant immunoreactions. By ABC method on the formalin-fixed, paraffin-embedded liver tissues, strong immunoreactivity was found in the periportal triad lesions and peripheral lesions of the hepatic lobules. Immunoreactive products showed diffuse fine granular in the cytoplasm of hepatocytes and sinusoidal cells. In some cells, immunoproducts marginate at the periphery of the cells. The intensive staining of the cytoplasm of infected cells allowed their exact differentiation from surrounding uninfected cells. The positive area involved coincided with histopathological lesion on serial liver sections. In conclusion, liver was proved to be a consistent target organ in RHD, and the immunoperoxidase method in the section of formalin-fixed, paraffin-embedded hepatic tissue could be broadly used for the routine diagnosis of the disease.

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Canine nervous-tissue tumors with features of peripheral nerve sheath tumor: histopathological and immunohistochemical findings (말초신경초 종양의 특징을 지닌 개 신경종의 조직병리학적 및 면역조직화학적 진단)

  • Lee, Sun-Gue;Lee, Jae-Ha;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.41 no.1
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    • pp.57-61
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    • 2018
  • Canine peripheral nerve sheath tumors (PNSTs) are spindle cell tumors that arise from Schwann cells, perineural cells, fibroblasts or all of them. Based on the morphology and biologic behavior, PNSTs are divided into benign PNST (BPNST) and malignant PNST (MPNST) forms. The aim of this study is to diagnose the two cases of neoplastic tissue samples with features of PNSTs by the histopathology and immunohistochemistry. The study was performed using two specimens from small animal clinic. The first case, A was a mass, 3~4 cm in diameter, extruded from vaginal mucosa of 10-year-old spayed female mixed-breed dog. And the second case, B was a subcutaneous mass, 1.5 cm in diameter, which is originated from right hind leg of 9-year-old castrated male mixed-breed dog. Two cases were stained with hematoxylin and eosin (H&E) for histopathological examination. And also immunohistochemistry (IHC) was performed by the avidin-biotin peroxidase complex (ABC) method with antibodies specific for the following proteins: S-100 protein, smooth muscle actin (SMA) and epidermal growth factor receptor (EGFR). In results, Antoni B schwannoma pattern characterized by pleomorphic, round and fusiform polygonal cells was seen in A. In B, Antoni A pattern, densely packed spindle cells arranged in interlacing bundles was seen in addition to Antoni B pattern. In IHC, cytoplasms of neoplastic cells were diffusely labeled for S-100 expression in A and B. For SMA, both A and B show negative expression. And for EGFR, A shows negative expression but B shows partially positive expression in areas of Antoni B schwannoma pattern. The histopathologic features of two cases coupled with the S-100 immunoreactivity led to a diagnosis of PNST. For SMA, both A and B show negative expression. The diagnosis of A will be a BPNST with the negative result and B will be a MPNST with the positive result for EGFR.

Studies on the pathogenesis of Korean isolate of Aujeszky's disease virus in experimentally infected piglets II. Immunohistochemistry and detection of viral nucleic acids by in situ hybridization (Aujeszky's disease virus 국내분리주 접종자돈의 병리발생에 관한 연구 II. 면역조직화학 및 in situ hybridization 기법을 이용한 항원과 핵산 검출)

  • Cho, Woo-young;Cho, Sung-whan;Park, Choi-gui;Kim, Jae-hoon;Hyun, Bang-hoon;Yoon, Yong-dhuk;Kweon, Chang-hee
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.859-871
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    • 1996
  • This study was conducted to elucidate the distribution of Aujeszky's disease viral nucleic acids and antigens in the central nervous system (CNS) of piglets. The first Korean isolate of Aujeszky's disease virus(ADV) that isolated from naturally infected piglets in Yang San, was inoculated into 32 day old piglets with $10^{5.9}TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at every 24hrs for 8 days. The immunohistochemistry (IHC) was conducted to detect the viral antigens in paraffin-embedded tissue sections using avidin-biotin-peroxidase complex (ABC) method. The viral nucleic acids were detected by in situ hybridization (ISH) using ADV specific DNA probe labeled with digoxigenin. The ADV antigens were detected in reticuloendothelial cells of spleen, lymph nodes and tonsil, alveolar walls, leptomeningeal vascular walls, inflammatory foci of each organ, and nerve cells. The viral nucleic acids were detected in the spinal trigeminal nucleus and its tracts of the pons and medulla oblongata by the ISH technique. The pathways of AD viruses in CNS were determined by IHC and ISH. In the intranasally inoculated group, the viruses in nasal mucosa moved to medulla oblongata and pons through the trigeminal nerve. In case of intramuscullarly inoculated group, viruses moved to brain via lymphoid organs or spinal nerves from sciatic nerves.

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Distribution of growth hormone-releasing factor- and somatostatin-immunoreactive neurons in the hypothalamus of the Korean squirrel(sciurus vulgalis coreae) (청서 시상하부의 growth hormone-releasing factor 및 somatostatin 면역반응신경세포의 분포)

  • Jeong, Young-gil;Son, Hwa-young;Yoon, Won-kee;Kim, Kil-soo;Won, Moo-ho;Ryu, Si-yun;Cho, Sung-whan;Kim, Moo-kang
    • Korean Journal of Veterinary Research
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    • v.35 no.4
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    • pp.671-681
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    • 1995
  • This study was attempted to investigate the topographical distribution, shape and immunoreactivity of growth hormone-releasing factor(GRF)- and somatostatin(SOM)-immunoreactive neurons in the hypothalamus of the Korean squirrels(Sciurus vulgalis coreae). For the light microscopical examination of immunohistochemistry, the brains were fixed with 4% paraformaldehyde solution by means of intracardiac perfusion. And the frozen sections($40{\mu}m$ thick) were stained immunohistochemically by ABC method. Distribution of GRF immunoreactive neurons($12-17{\mu}m$) was highest in the paraventricular nucleus, moderate in the periventricular and supraoptic nuclei, and low in the arcuate nucleus and lateral hypothalamic area. Their immunoreactive fibers were found very high in the median eminence, moderately in the supraoptic, paraventricular and periventricular nuclei, and low in the arcuate nucleus and lateral hypothalamic area. SOM immunoreactive perikarya($14-18{\mu}m$) were found moderately in the periventricular nucleus near the subependymal layer of the third ventricle, and low in the arcuate and suprachiasmatic nuclei. SOM immunoreactive fibers were found high in the median eminence, and moderately or low in the arcuate and periventricular nuclei.

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Regional distribution and relative frequency of the gastrin, secretin and pancreatic polypeptide-immunoreactive cells in the gastrointestinal tract of the fetus of Korean native goat (한국재래산양 태자의 위장관에 있어서 gastrin, secretin 및 pancreatic polypeptide 면역반응세포의 분포 및 출현빈도에 관한 연구)

  • Lee, Hyeung-sik;Ku, Sae-kwang;Lee, Jae-hyun
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.12-19
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    • 1999
  • The regional distributions and relative frequencies of the gastrin, secretin and pancreatic polypeptide(PP)-immunoreactive cells in the gastrointestinal tract of the fetus(180 days of gestation) of Korean native goat were studied with immunohistochemical(ABC) methods. Gastrin-immunoreactive cells were detected in fundus, pylorus and duodenum and these cells were most predominant in pylorus. Secretin-immunoreactive cells were observed in pylorus, duodenum and ileum. PP-immunoreactive cells were restricted to fundus. These immunoreactive cells were situated in surface epithelium and mucosal gland regions. The regional distribution and relative frequency of PP-immunoreactive cells was somewhat different to the adult Korean native goat. Immunoreactive cells in the surface epithelial regions were open typed cells which were spindle shaped cells but closed typed cells which were round or/to spherical shaped cells were observed in the mucosal gland regions.

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GFAP IMMUNOREACTIVITY IN TRIGEMINAL GANGLION SATELLITE CELLS AFTER PULP EXPOSURE IN RAT (흰쥐에서 치수노출 후 삼차신경절의 신경절아교세포에서 GFAP-IR의 변화)

  • Kim, Heung-Jung;Moon, Joo-Hoon
    • Restorative Dentistry and Endodontics
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    • v.22 no.2
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    • pp.782-791
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    • 1997
  • Glial fibrillary acidic protein(GFAP) are a group of intermediate filaments that are distributed in the cytoplasm of many type of glial cells. The purpose of this study was to determine change of GFAP immunoreactivity(GFAP-IR) in rat trigeminal ganglion satellite cells in response to pulp exposure. The immunohistochemistry was carried out using the avidinbiotin-peroxidase complex(ABC) method and subsequently stained with AEC(3-aminoethyl-9-carbasol). 1. Contol group; Central root astrocytes had strong GFAP-IR, but ganglion satellite cells occasionlly had GFAP-IR. This reaction patterns of ganglion satellite cells was not concenturated in any specific region of trigeminal ganglion. 2. Three day pulp exposure group; There was a highly GFAP-IR in satellite cells of trigeminal ganglion in maxillary region. GFAP-IR in neighboring mandibular and ophthalmic regions was less intense compared to maxillary region. 3. Seven day pulp exposure group; In this group, GFAP-IR that was increased compared to control group was seen in the maxillary region. But GFAP-IR was less intense compared to three day pulp exposure group. These results suggest that GFAP in satellite cell increase in specific region of trigeminal ganglion after pulp exposure and offer useful tool in trigeminal pain research.

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GFAP IMMUNOREACTIVITY IN SATELLITE CEllS OF TRIGEMINAL GANGLION FOllOWING AXOTOMY OF INFERIOR ALVEOLAR NERVE IN RAT (흰쥐에서 하치조신경 절단에 따른 삼차신경절 위성페포에서 GFAP-IR의 변화)

  • Lee, Chang-Seop;Lee, Sang-Ho;Kim, Heung-Jung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.25 no.1
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    • pp.249-256
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    • 1998
  • Glial fibrillary acidic proteins (GFAP) are a group of intermediate filaments that are distributed in the cytoplasm of glial cells. GFAP immunoreactivity (GFAP-IR) increase after central and peripheral nerve injuries. The purpose of this study was to determine change of GFAP-IR in rat trigeminal ganglion satellite cells following the axotomy of inferior alveolar nerve(IAN). The immunohistochemistry was carried out using the avidin-biotin-peroxidase complex(ABC) method. 1. Control group : Astrocytes in central root of trigeminal ganglion had strong GFAP-IR, but satellite cells of trigeminal ganglion occasionally had GFAP-IR. The patterns of reactivity in satellite cells of trigeminal ganglion were not concenturated in any specific region of trigeminal ganglion. 2. Three day group after IAN axotomy : There were highly GFAP-IR in satellite cells of trigeminal ganglion in mandibular region. GFAP-IR in maxillary and ophthalmic regions were less intense compared to mandibular region. 3. Seven day group after IAN axotomy : GFAP-IR that were increased compared to control group were seen in the mandibular region. But GFAP-IR were less intense compared to three day group. These results suggest that GFAP-IR increase in specific region of trigeminal ganglion following peripheral axotomy. therefore we suppose that GFAP study offer research tool in trigeminal neuralgia.

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