• 대한수의학회지
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• 제29권4호
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• pp.541-548
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• 1989

오제스키병 바이러스를 배양세포에다 감염시켜, 냉동 및 araldite포매 초박절편에서 protein A-gold labeling을 통해 바이러스항원 검출을 시도하였다. 오제스키병 바이러스항원은 10nm gold probes로 표지되었으며, 전자밀도가 높은 gold 입자들은 바이러스의 nucleocapsid와 envelope에서 주로 관찰되었고, 초냉동박절표본에서의 immunogold labeling은 조직구조물들과 극히 미미한 비특이결합만을 보였다. 초냉동박절표본에서의 immunogold labeling은 오제스키병 바이러스항원을 검출하는데 있어 효과적이었으며, 이는 또한 여러가지 바이러스들과 속주세포들간의 상호작용에 관한 면역세포화학적 연구에 크게 이용될 수 있을 것으로 생각된다.

• 대한수의학회지
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• 제28권2호
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• pp.365-369
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• 1988

본 연구는 감염돼지와 조직배양세포의 동결 및 파라핀 절편에서 immunogold-silver(IGS)법을 이용하여 오제스키병 바이러스를 검출코자 하였으며, 이를 위해 자돈 5두와 돼지뇌유래 섬유 아세포 및 BHK 세포를 공시하였고, IGS법과 immunoperoxidase법의 1차 항원으로는 2종의 단크론성 항체와 고도면역혈청을 사용하였다. IGS법은 양성부위에서 흑색의 미세한 과립의 침착을 볼 수 있었고, immunoperoxidase법에 비해 탁월한 효과를 얻었으며, IGS법은 조직내에 존재하는 각종 바이러스 항원을 검출하는데 이용할 수 있을 것으로 생각된다.

• International Journal of Oral Biology
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• 제40권4호
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• pp.183-187
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• 2015

The present study was aimed to evaluate the influence of glutaraldehyde (GA) concentration on multiple electron microscopic (EM) immunostaining using pre-embedding peroxidase and post-embedding immunogold method. Influence of various concentrations of GA included in the fixative on immuoreactivity was assessed in the multiple immunostaining using antisera against anti-transient receptor potential vanilloid 1 (TRPV1) for peroxidase staining and anti-GABA for immunogold labeling in the rat trigeminal caudal nucleus. Anti-TRPV1 antiserum had specificity in pre-embedding peroxidase staining when tissues were fixed with fixative containing paraformaldehyde (PFA) alone. Immunoreactivity for TRPV1 was specific in tissues fixed with fixative containing 0.5% GA at both perfusion and postfixation steps, though the immunoreactivity was weaker than in tissues fixed with fixative containing PFA alone. Tissues fixed with fixative containing 0.5% GA at the perfusion and postfixation steps showed specific immunogold staining for GABA. The results of the present study indicate that GA concentration is critical for immunoreactivity to antigens such as TRPV1 and GABA. This study also suggests that the appropriate GA concentration is 0.5% for multiple immunostaining with peroxidase labeling for TRPV1 and immunogold labeling for GABA.

• Applied Microscopy
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• 제30권4호
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• pp.403-412
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• 2000

1. 아메바 항-액틴의 Ag-Ab 반응과 이를 항-생쥐 IgG-금 입자로 표지한 결과는 주로 정세 포 및 정자의 핵질에 특이적으로 표지되었고 첨체에서는 그 반응을 볼 수 없었다. 2. 표지된 항-액탠 금 입자로 볼 때 정자 핵질의 G-액틴 또는 G-actin oligomer는 정세포의 F-액틴에서 유래되는 것으로 사료된다. 3. 첨체의 액틴은 주로 F-액틴으로 첨체돌기 형성에 참여하지 않는 위상인 것으로 관찰된다. 4. 미세구조의 변화, 정세포 체적의 감소, 정세포 및 정자 핵에 표지되는 금 입자의 증가와 이들 현상이 나타나는 동시성으로 볼 때 정세포 핵의 형태변화의 내용은 응축이고 이는 F-actin의 탈중합 반응에 의한 G-액틴의 생성이 원인일 것으로 사료된다.

• 대한수의학회지
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• 제35권3호
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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.

• 한국패류학회지
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• 제15권2호
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• pp.81-92
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• 1999

The histochemical, cytochemical, and immunocytochemical investigations were conducted to find out the cellulase activity in the accessory glands of the digestive system of the oriental land snail Nesiohelix samarangae under the LM, SEM, and TEM. The cellulase activity was shown in the epithelium of th digestive gland by labelling with the immunogold (protein-A gold) particles. The epithelial cells showing the cellulase activity were Type 1 and Type 3 cells out of five types of the epithelial cells of the digestive gland. None of epithelial cells of the mucus gland and the salivary gland and the salivary gland were not labeled with the immunogold particles.

• 한국미생물·생명공학회지
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• 제17권5호
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• pp.499-503
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• 1989

Cryptosporidium이 인체 내에서 복잡한 life cycle를 거치면서 각 stage마다 만들어지는 단백질 항원의 관계성에 관한 보고가 없어 Cryptosporldiosis에 대한 치료제 개발에 어려움이 있었다. 본 연구는 단일군 항체와 Immunogold labeling 기술을 이용하여 주요 extracellular stages인 sporozoites와 merozoites의 antigenic relatedness를 살펴보았다. BALB/C 쥐로부터 merozoites에 대한 단일군 항체(Jo3)를 분리하였으며 IgG3형이었다. 정제된 sporozoites를 SDS-PAGE로 분리한 후 Western blot을 이용하여 Jo3를 반응시킨 결과 3,500Daltons 크기의 sporozoites 항원을 인식하였다. Jo3를 Cryptosporidium에 감염된 tissue section에 반응시킨 후 Immunoelectron microscopy를 이용하여 3.5-kDa 항원의 위치와 sporozoites와 merozoites가 똑같은 단백질 항원을 만드는가를 추적해 본 결과 3.5-kDa 단백질 항원이 두 stages에서 공동으로 합성되는 것으로 나타났으며 이 항원은 표면과 세포질 내에 위치하고 있었다.

• Parasites, Hosts and Diseases
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• 제34권4호
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• pp.215-224
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• 1996

작은와포자충의 세포골격 단백질 분포를 알아보기 위하여 actin, tropomyosin, $\alpha$-actinin 및 troponin-T의 분포를 이중면역 황금표지법 (double immunogold labeling method)으로 관찰하 였다 관찰된 모든 발육 단계의 충체에서 매우 많은 양의 tropomyosin이 세포질 및 세포막에 분포하고 있음이 밝혀졌으며 actin보다도 더 많은 양이 관찰되었다. ${\alpha}-actinin$의 경우는 주로 세포막에 소량이 분포하였으며 Troponin-T는 어느 발육 단계에서도 관찰되지 않았다 이 연구의 결과 tropomyosin의 분포 정도도 미루어 볼 때 이 단백질이 작은와포자충에서 매우 중요한 역할을 수행할 것으로 추측되며, 주로 세포막에 다량으로 분포하는 actin, tropomyosin 및 ${\alpha}-actinin$은 면역진단 및 면역치료의 대상으로 이용할 가능성이 있을 것으로 생각된다.

• Applied Microscopy
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• 제25권3호
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• pp.1-19
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• 1995

The present study was performed to clarify the effect of vagus nerve stimulation on the enterochromaffin(EC) cells in the body of the stomach, the first part of the duodenum and the ceceum of rats by using routine electron microscopy and immunogold labelling. The changes in the ultrastructure and in the labelling density of the gold particles of the EC cells were investigated after vagus nerve stimulation. The vagus nerve was electrically stimulated with a square wave pulse generator for a duration of 5 minutes each, a total of 8 times at 2 minute intervals. Immunogold labelling demonstrated that the epithelial serotonin immunoreactive cells of the gastrointestinal tract are EC cells containing characteristic pleomorphic granules. Immunocytochemically labelled gold particles were largely concentrated in the dense matrix of the granules of the EC cell, and the labelling density of the gold particles considerably increased after the vagus nerve stimulation. Except for a slight activation of Golgi complexes, no remarkable changes in the ultrastructures of the EC cells were noted after the vagus nerve stimulation. The above results suggest that vagus nerve stimulation may activate serotonin biosynthesis in EC cells.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.468-473
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• 1996

The genome of tomato spotted wilt virus (TSWV) is composed of three RNA segments, S, M, and L RNA and the 5.0 kb M RNA encodes two glycoproteins Gl, G2 and NSm protein of unknown function. In an effort to investigate the function of the NSm protein, antibody was raised against NSm fusion protein overexpressed in Escherichia coli. This antibody was used to detect the NSm protein by using western blot analysis and electron microscopic observation after immunogold labelling. For the cloning of the NSm gene, total RNA extracted from a TSWV infected plant was used for cDNA synthesis and polymerase chain reaction (PCR) instead of going through time-consuming virus purification. A protein band specifically reacting to the NSm antibody was detected from TSWV inoculated plants. The NSm protein was detected in the cell wall fraction and in pellet from low speed centrifugation when the infected plant tissue was fractionated into 4 fractions. In the immuno-electron microscopic observation, gold particles were found around the plasmodesmata of infected plant tissue. These results suggest that the NSm protein of TSWV plays some role in cell-to-cell movement of this virus.