• 제목/요약/키워드: Immunogold labeling

검색결과 28건 처리시간 0.021초

Immunoelectron Microscopic Localization and Analysis of Herpes simplex Virus Type 1 Antigens

  • Chung, Charles C.;Lee, Hyung-Hoan;Cho, Myung-Hwan
    • Journal of Microbiology and Biotechnology
    • /
    • 제10권5호
    • /
    • pp.714-720
    • /
    • 2000
  • Antigens of Herpes simplex virus type 1 (HSV-1) strain F were immunoblotted to identify the most immunodominant one, and the localization of this antigen was then studied using immunoelectron microscopy. The 67.8 kDa antigen appeared to be the most immunodominant one in a mouse model, and it showed randomly scattered and partially clustered distribution on the surface of the virion. The localization study was performed using immunogold with polyclonal anti-HSV-1 sera produced from BALB/c mice, and immunofluorescence demonstrated that the viral products in the HSV-2 infected Vero cells were distributed throughout the infected host cell, however, mainly on the surface of the host membrane.

  • PDF

면역황금 표기법을 이용한 간흡충의 체액 항원에 관한 연구 (A study on the body fluid antigen of Clonorchis sinensis using immunogold labeling method)

  • 주봉덕;임한종;김수진
    • Parasites, Hosts and Diseases
    • /
    • 제28권1호
    • /
    • pp.11-24
    • /
    • 1990
  • 간흡충의 항원성 부위를 규명하기 위하여 간흡충 충체로부터 분리한 체액을 실험 토끼에 주사하여 면역시킨 IgG와 간흡충에 감염된 실험 토끼의 IgG를 간흡충 조직 항원에 반응시키고 면역황금 표식법을 이용하여 전자현미경으로 관찰한 바 그 결과는 다음과 같다. 간흡충의 조직세포 사이의 구성물인 세포간질과 맹관의 내용물, 그리고 맹관 막 구조물은 충체에 감염된 IgG와 체액 항원에 면역된 IgG 모두에서 항원-항체 반응이 관찰되었지만 체액 항원으로 면역된 IgG에서 다소 강한 반응이 나타났다. 표피의 기저층과 난황선의 난황 물질은 충체에 감염된 IgG에서는 반응이 나타나지 않았으나 체액 항원에 면역된 IgG에서는 강한 반응이 나타나 이들을 구 성하는 물질은 체액 반응 IgG에 대한 특이 항원으로 생각되었다. 배설관의 내용물과 막 구조물은 충체에 감염된 IgG에 대한 반응이 특이하였으나 체액 항원에 면역된 IgG에 대한 반응은 매우 미 약 하여 항원성에 차이점이 있는 것으로 생각되었다. 수정낭의 정자세포 간질은 충체 감염 IgG에 반응 하고 정자세포의 두부는 체액 항원으로 면역된 IgG에 반응하여 매우 상이한 항원성이 나타났다. 이상의 결과로 보아 충체의 세포간질과 맹관 구성물은 충체 감염시와 체액 항원에 면역된 IgG에 대하여 반응의 정도 차이는 있으나 항원성으로 관찰되는 것은 동일하였다. 그러나 수정낭의 세포간질과 배설낭의 구성물질은 충체에 감염된 IgG에 대한 특이 항원이며 수정낭 내의 정자들의 두부와 난황을 구성하는 물질은 체액 면역 IgG에 대한 특이 항원인 것으로 생각된다.

  • PDF

한국 연근해산 두족류 (Todarodes pacificus and Octopus minor) 시엽내 Dopamine 및 $Calbindin-D_{28K}$의 분포에 관한 면역전자현미경적 연구 (Immuno-Electron Microscopic Studies on the Distribution of Dopamine and $Calbindin-D_{28K}$ in the Optic lobes of Cephalopods (Todarodes pacificus and Octopus minor) inhabiting the Korean waters)

  • 한종민;장남섭
    • Applied Microscopy
    • /
    • 제32권2호
    • /
    • pp.175-183
    • /
    • 2002
  • 한국 연근해산 살오징어와 서해낙지의 시엽에서 신경전달물질을 분비하는 neuron의 특성 및 기능을 확인하기 위해 rabbit anti-dopamine (TH)과 rabbit anti-calbindin-$D_{28K}$ 등의 항체를 이용한 면역염색과 면역금지법을 시행한 결과는 다음과 같았다. 항-dopamine을 사용한 면역염색 결과 살오징어에서는 외과립세포층 상단에 위치한 대형 무축삭세포와 수질부의 섬을 이루는 세포들 중 $2{\sim}3$개의 비교적 소량의 세포만이 양성반응을 보인 반면, 서해낙지에서는 외과립세포층 상단 및 중간 부위의 $2{\sim}3$개의 세포와 수질부섬을 이루는 5개 이상의 세포에서 강한 양성반응을 보였다. 항-calbindin-D28k인 경우는 살오징어의 외과립세포층 상단에 위치한 $2{\sim}3$개의 소형 무축삭세포와 내과립세포층 하단에 위치한 $1{\sim}2$개의 세포에서 양성반응을 보인 반면, 서해낙지인 경우는 외과립세포층에서 4개 정도의 세포에서 양성 반응을 보였으나, 내과립세포층에서는 반응을 확인할 수 없었다. 면역금지법을 시행한 결과 살오징어에서는 항-dopamine과 항-calbindin에 면역반응을 보인 세포들의 세포질 $0.5{\mu}m^2$$17{\sim}26$개 정도의 비교적 많은 수의 금입자가 표지된 반면, 서해낙지에서는 세포질 $0.5{\mu}m^2$당 평균 10개 정도인 소량의 금입자만이 표지되어, 살오징어가 서해낙지에 비해 표지된 금입자가 거의 2배 정도 많았다.

Ultrastructural Localization of Cryptosporidium parvum Antigen Using Human Patients Sera

  • Lee, Jong-Gyu;Han, Eun-Taek;Park, Woo-Yoon;Yu, Jae-Ran
    • Parasites, Hosts and Diseases
    • /
    • 제47권2호
    • /
    • pp.171-174
    • /
    • 2009
  • The antigen location of Cryptosporidium parvum, which stimulates antibody formation in humans and animals, was investigated using infected human sera. Immuno-electron microscopy revealed that antigenicity-inducing humoral immunity was located at various developmental stages of parasites, including asexual, sexual stages, and oocysts. The amount of antigen-stimulating IgG antibodies was particularly high on the oocyst wall. The sporozoite surface was shown to give stimulation on IgG and IgM antibody formation. Trophozoites implicated the lowest antigenicity to humoral immunity, both IgG and IgM, by showing the least amount of gold labeling. Immunogold labeling also provided clues that antigens were presented to the host-cell cytoplasm via feeder organelles and host-parasite junctions.

Application on Microwave Energy in the Preparation of Fish Samples for Electron Microscopic Observation

  • Kim Soo Jin;Oh Hae Keun;Song Young-Hwan;Chung Hyun-Do;Kim Young-Tae;Park Nam-Kyu;Choi Tae-Jin
    • Fisheries and Aquatic Sciences
    • /
    • 제1권2호
    • /
    • pp.187-191
    • /
    • 1998
  • Chemotherapy can not be applied for the control of fish viral diseases because viruses depend on host machinery for their replication. Although new control strategies including vaccination are under development, avoidance of virus introduction by rapid and correct diagnosis is the best way of fish viral disease control. Although observation of virus particles with an electron microscope is an easy method for virus detection, it take a few days for the sample preparation. In order to shorten the sample preparation time, microwave radiation was applied in the procedure. With this method, 15 seconds was enough for fixation of virus infected fish samples or cultured cells inoculated with infectious hematopoietic necrosis virus, which takes 2-4 hours with routine methods. Also four minutes was enough for polymerization of embedding resin which takes 24-48 hours with routine methods. Samples prepared with microwave were good enough for direct electron microscopic observation and immunogold labeling assay.

  • PDF

Distribution of Hemicelluloses in Warts and the Warty Layer in Normal and Compression Wood Tracheids of Cryptomeria Japonica

  • Kim, Jong-Sik;Awano, Tatsuya;Yoshinaga, Arata;Takabe, Keiji
    • Journal of the Korean Wood Science and Technology
    • /
    • 제39권5호
    • /
    • pp.420-428
    • /
    • 2011
  • The distribution of arabino-4-O-methylglucuronoxylans (AGXs) and O-acetyl-galactoglucomannans (GGMs) in warts and the warty layer of tracheids in normal wood (NW) and compression wood (CW) of Cryptomeria japonica was investigated. Under field emission scanning electron microscope (FE-SEM) observation, warts and the warty layer of delignified NW and CW tracheids were degraded by xylanase treatment, indicating that warts and the warty layer contain high amounts of AGXs. However, the effect of xylanase was not observed in NW and CW tracheids before delignification, suggesting that AGXs in warts and the warty layer may be encrusted with lignin. After ${\beta}$-mannanase treatment, no noticeable changes were observed in warts and the warty layer of NW tracheids, indicating that warts and the warty layer contain either no or very few GGMs. Similar results to FE-SEM observations were also observed with immunogold labeling. AGX labeling was observed in warts and the warty layer of NW and CW tracheids, while GGM labeling was not detected. NW tracheids showed a much stronger density of AGX labeling than did CW tracheids in warts and the warty layer, indicating differences in the chemical compositions of warts and the warty layer between NW and CW tracheids.

Localization of Single Chain Fv Antibodies (scFv) in Transgenic Tobacco Ptants Showing Resistance against Tomato Bushy Stunt Virus

  • Jeun, Y.C.;Boonrod, K.;Nagy, P.;Conrad, U.;Krczal, G.
    • 한국식물병리학회:학술대회논문집
    • /
    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
    • /
    • pp.75.2-75
    • /
    • 2003
  • To develop an effective protection strategy against tomato bushy stunt virus (TBSV), tobacco plants expressing single-chain Fv antibodies (scFv), were established. A previous had shown that the replication activity of viral replicase was inhibited by the selected scFvs. Moreover, no systemic symptom was found after virus inoculation on leaves of wt N. benthamiana infiltrated with an Agrobacterium suspension resulting i3l expression of the scFvs. However, control plants showed systemic symptoms. In this study the localization of the scFvs within two transgenic plant lines, (CP28H3, CP-P55) was demonstrated using immunogold labelling. The gold particles, indicating the presence of scFv, were mostly found In the cytoplasm of the plant cells including chloroplasts and in the cell walls. However, they were hardly found in the vacuole, nucleoplasm and intercellular spaces. Gold particles often accumulated in either the cytosol or chloroplasts showing a specific labeling, There was no difference in type of gold labeling between both transgenic lines. The localization of the scFv in the cytoplasm further conforms the inhibition of the RNA-dependent RNA polymerase (RdRp) by the selected scFv because it is known that the RdRp is localized to membraneous cytosolic structures.

  • PDF

미토콘드리아 크리스테에 존재하는 cytochrome-c-oxidase의 단백질 소단위 분포에 관한 연구 (A Study on the Distribution of Cytochrome-c-oxidase Subunit in the Cristae of Mitochondria)

  • 김수진;이지현;정차권
    • Applied Microscopy
    • /
    • 제24권4호
    • /
    • pp.41-51
    • /
    • 1994
  • The topology of the enzyme has been investigated by biochemical studies including chemical labeling and cross linking. Thirteen subunits(polypeptides) of the cytochrome-c-oxidase have localistic characteristics of existing in the matrix side or cytoplasmic side in the mitochondria. In order to observe the distribution of the enzyme subunit on the mitochondria membrane, immunogold-labeling methods were employed. Antibody was obtained from the serum of immunized rabbit with enzyme subunit antigen which was obtained from cytochrome-c-oxidase of the beef heart muscle mitochondria. Beef heart muscle tissue as a tissue antigen was stained with immunized rabbit IgG and protein A gold complex. Electron microscopy has identified the existance of cytochrome-c-oxidase subunit $Mt_I,\;Mt_{II}\;and\;Mt_{III}$ on the membrane of cristae and outer chamber of mitochondria and the subunit $C_{IV}$ on the membrane of cristae and matrix of mitochondria. Particularly, the subunit $C_{IV}$ was also observed to exist in the sarcoplasm of muscle tissue.

  • PDF

Binding of Tp92 homolog of Treponema denticola to fibronectin and epithelial cells

  • Jun, Hye-Kyoung;Lee, Sung-Hoon;Lee, Hae-Ri;Choi, Bong-Kyu
    • International Journal of Oral Biology
    • /
    • 제33권2호
    • /
    • pp.45-50
    • /
    • 2008
  • Treponema denticola is the best studied oral spirochete and numerous studies have shown that it is strongly associated with periodontitis and expresses several putative virulence factors. In this study, we report on a surface protein of T. denticola, Td92, which is homologous to Tp92 of Treponema pallidum, an agent of syphilis. Immunofluorescence assay and immunogold labeling with anti-Td92 Ab revealed that Td92 had surface-exposed epitopes. And Td92 was capable of binding to fibronectin and KB cells, an oral epithelial cell line. In addition, Td92 could enter the KB cells. These results indicate that Td92 is a fibronectin-binding protein which can bind to and internalize into the host cells, facilitating the virulence of T. denticola.