• Title/Summary/Keyword: Immunocytochemical

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The Principles and Practices of Immunocytochemical method in Light Microscopic Level (광학 현미경적 수준에서의 면역조직화학적 방법의 원리 및 실제)

  • Kim Jin-Sang
    • The Journal of Korean Physical Therapy
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    • v.3 no.1
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    • pp.229-250
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    • 1991
  • The study was carried out to investigate and review the principles and practices of immunocytochemical method in light microscopic level. The results were as follows. 1. Immunocytochemistry is the method to search out the intracellular position of the specific meterials using antigen -antibody reaction. 2. The chief items in immunocytochemistry are antigen, antibody and chromogen. 3. The identifical fixation is cardiac perfusion fixation. 4. The tissue slides must be prepared by vibratomy. 5. All stainings are carried out with free floating staining method. 6. There are polyclonal and monoclonal antibodies used in immunocytochemistry.

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Distribution of Substance P Immunoreactive Neurons and Their Synaptic Organization in the Cat Thoracic Cord (고양이 흉수에서 Substance P 면역반응 신경원의 분포와 연접연구)

  • Lee, Seung-Kyun;Park, Soo-Seog
    • The Korean Journal of Pain
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    • v.9 no.2
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    • pp.326-335
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    • 1996
  • Background: Though a number of studies have described the distribution of substance P(SP)-like immunoreactivity in the spinal cord, they have been focused on lamina I and II of the dorsal horn and there are little morphological studies on the topographic distribution and ultrastructure of the SP immunoreactive neurons especially in the ventral horn of the spinal cord. this study was conducted to identify distribution pattern of SP immunoreactive neurons and to difine the synaptic organization of their processes in ventral horn of the thoracic cord of the cat by preembbeding immunocytochemical method using SP antiserum. Methods: Five adults cats of either sex were used and deeply anesthetized by intramuscular injection of ketamine. After removal of the spinal cord, samples of thoracic cord were taken and placed in fresh fixative at $4^{\circ}C$ for 2 hours. Transverse sections $50{\mu}m$ thick were processed using the preembbeding immunocytochemical method and incubated consecutively in the specific primary antibody and the 10% normal goat serum, the rabbit anti-substance P antiserum, the biotin-labelled goat anti-rabbit IgG and finally the avidin-biotin-peroxidase complex. The processed tissue sections were throughly washed and stained in the black with 1% uranyl acetate. Section were examined on a electron microscope. Results: 1) SP immunoreactive neurons were observed in the gray matter around central canal. 2) In lamina I and II SP immunoreactivity was observed in both myelinated and unmyelinated nerve fibers, but in ventral horn only in the unmyelinated nerve fibers. 3) SP immunoreactive axon terminals with small round and large dense core vesicles made chemical synapses onto the dendrites of motor neurons in the ventral horn. Conclusion: SP immunoreactive neurons might play an important role in modulation of motor neurons in the ventral horn of the thoracic cord of the cat.

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Cytochemical and Immunocytochemical Study on the Cellulase Activity in the Digestive Tract of the Land Snail Nesiohelix (동양달팽이 Nesiohelix samarangae 소화관에서의 cellulase 활성에 대한 세포화학적 및 면역세포화학적 연구)

  • 정계헌;이용석;김은정
    • The Korean Journal of Malacology
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    • v.14 no.2
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    • pp.149-159
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    • 1998
  • In order to observe the anticellulolytic localization in the epithelia of the digestive tract such as esophagus, crop, and intestine of a Korean land snail N. samarangae, a cytochemical method and a immunogold labelling method were applied. For the cytochemical study on the cellulase activity, Benedict reaction method applied. And for the immunocytochemical study, the rabbit serum immunoglobuins (IgG) was obtained from the rabbits injected with cellulase which was extracted from body fluid of the snail. The digestive tract tissues of N. samarangae were fixed with 4% paraformaldehyde and 2% OsO4 and embedded in Lowicryl K4M at -40$^{\circ}C$ under UV light (360 nm). The thin sections were loaded on the nickel grids and stained with the serum IgG and protein A-gold complex (particle size: 10 nm). Observations were undertaken with transmission electron microscope (Jeol, JEM-1010). The epithelium of the digestive tract was consisted of five types of cells. In the cytochemical study, the reaction products were found along the periphery of the vacuoles derived from the Bebedict reaction. In the immunocytochemical study, the protein-A gold particles were selectively labelled in Type 1, Type 3 and Type 4 cells in intestinal tissue. membranes of rER, in the surrounding cytoplasm of the rER and secretory granules, and in the apical cytoplasm of the cells. On the material being secreted from the apical cytoplasm was also labelled with the immunogold particles. The all results obtained throughtout present study suggest that the intestinal epithelium of the snail N. samarangae seretes cellulase as one of digestive enzymes.

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Fine Needle Aspiration Cytology of Subacute Granulomatous Thyroiditis -A Clinico-Cytological Review of 10 Cases with Immunocytochemical Analysis- (아급성 육아종성 갑상샘염의 세침흡인 세포소견 -아급성 육아종성 갑상샘염 10예의 임상-세포소견 및 면역세포염색 소견 분석-)

  • Kim, Do-Kyung;Pyo, Ju-Yeon;Park, Jong-Pil;Jin, Lian-Hua;Jeong, Woo-Hee;Son, Eun-Ju;Hong, Soon-Won
    • The Korean Journal of Cytopathology
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    • v.19 no.1
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    • pp.27-33
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    • 2008
  • Although subacute granulomatous thyroiditis(SGT) is usually diagnosed clinically, papillary carcinoma or other thyroid conditions must be considered in the differential diagnosis. We retrospectively reviewed the clinical and fine-needle aspiration(FNA) cytologic findings seen in 10 SGT cases to decide what are the most reliable cytologic findings and the most helpful molecular tools for reaching a confident cytologic diagnosis. The most representative smear slides were retrieved to perform immunocytochemistry for cytokeratin 19(CK19) and Ret protein. Five papillary carcinomas(PTCs) were included as controls. The constant and typical cytologic findings of SGT were multinucleated giant cells(MGCs) (100%), epithelioid granulomas(90%), an inflammatory dirty background(90%) and plump transformed follicular cells(80%) without fire-flare cells, oncocytic cells or transformed lymphocytes. The immunoreactivities for CK19(37.5%) and Ret(10%) of the follicular cells of SGT were less than those(CK19 and Ret:100%) of PTC. CK19 immunoreactivity of the MGCs was seen in only one case of PTC. There was no significant difference between CK19 and Ret immunocytochemical staining for the MGCs of both SGT and PTC. The results of this study demonstrate that the cytological diagnosis of SGT can be improved by employing a combination of the typical and constant diagnostic cytological features and immunocytochemical results.

Immunocytochemical Study for Lactalbumin in Alveolar Macrophage of Human Milk Aspirated Mouse (인유(人乳) 흡인 백서의 폐포 대식세포에서 Lactalbumin에 대한 면역세포화학적 연구)

  • Han, Byoung Kil;Chung, Young Hun
    • Clinical and Experimental Pediatrics
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    • v.46 no.6
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    • pp.536-540
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    • 2003
  • Purpose : Aspiration of foreign material into the lungs can cause acute or chronic pulmonary diseases. It is difficult to detect small amounts of aspiration due to the lack of safe, sensitive and specific diagnostic tests. Recently, in animal or human studies, it has been reported that immunochemistry for lactalbumin can be used to detect the minimal aspiration. So, the authors' investigation was designed to determine whether human milk phagocytized alveolar macrophages can be detected in human milk aspirated mice. Methods : Sixty four male mice, 6-8 weeks old and 30-40 gm weighing, were used for this study. About 0.05 mL of human milk or normal saline were given intranasally once per day for 1 day or 3 days. Under anesthesia with ketamine and xylazine, the trachea of each mouse was cannulated with an 18G Jelco needle and then, each mouse's lungs were lavaged three times with 0.5 mL of phosphate buffer solution at 2, 8, 24, and 48 hours after the last milk or normal saline instillation. Cells in bronchoalveolar lavage fluid were stained with Oil Red O and immunocytochemistry for alpha-lactalbumin. Results : Immunocytochemical reactivity for alpha-lactalbumin or lipid-laden alveolar macrophages were not observed in the normal saline aspirated groups. Immunocytochemical reactivity for alpha-lactalbumin were observed in the human milk aspirated groups. They showed a peak at 8 hours and decreased markedly at 24 hours but persisted even at 48 hours after aspiration. Immunocytochemical stain positive alveolar macrophages were noted similarly in number between single and multiple aspiration groups. Conclusion : These observations suggested that alveolar macrophages for lactalbumin could be more easily detected on immunocytochemistry than Oil Red O stain, and immunocytochemistry could be used as a sensitive and specific diagnostic test for the detection of human milk aspiration.

Improvement of Neuronal Differentiation by PDE4 Inhibition in Human Bone Marrow-mesenchymal Stem Cells (인간 골수유래-중간엽 줄기세포(hBM-MSCs)에서 PDE4 억제조절을 통한 신경세포 분화 효율 개선)

  • Jeong, Da Hee;Joe, I-Seul;Cho, Goang-Won
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1355-1359
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    • 2016
  • Human bone marrow mesenchymal stem cells (hBM-MSCs) can differentiate into various cell types including osteoblasts, adipocytes, chondrocytes, and myocytes. Previous studies, including our own, have shown that MSCs can also differentiate into neuron-like cells. However, their rate of neuronal differentiation is not sufficient for application to stem cell therapy, which requires well-defined cell types. For this purpose, we first examined the expression of neuronal lineage markers (GFAP, MAP-2, KCNH1, Nestin, NF-M, and Tuj-1) by real-time PCR, western blot, and immunocytochemical staining. The expressions of the astrocyte marker GFAP and neuronal markers NF-M and Tuj-1 increased in neuronal differentiated MSCs (dMSCs). To improve the neuronal differentiation efficiency, PDE4, an important signaling intermediator in the progression of neuronal differentiation, was modulated using well-known inhibitors such as rolipram or resveratrol and then differentiated into neuronal cells (Roli- or RSV-dMSCs). The expressions of NF-M, Tuj-1 were increased while that of GFAP decreased in Roli- and RSV-dMSCs, which were examined by real-time PCR, western blot, and immunocytochemical staining. From these experiments, we have found that the neuronal differentiation efficiency can be ameliorated by the modulation of PDE4 activity.