• Biomedical Science Letters
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• v.5 no.2
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• pp.155-165
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• 1999

This study involved applying rat sera (control, infected and immunized) to adult worm tissue in order to measure the antigenic response. A serum was obtained from rats infected with E. hortense metacercaria for 4 weeks. Another immunized serum was taken from rats given a muscle injection with crude adult worm antigen. The detection of antigenic response in E. hortense tissue was made by immunogold labeling method and measured through gold particles impregnated in the tissue. The antigenic sites, those with the highest density of gold particles, were the tegmental syncytium, vitelline cells, seminal receptacle and cecum.

• Parasites, Hosts and Diseases
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• v.29 no.1
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• pp.31-42
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• 1991

In order to observe the antigenic localization in the tissues of the young adult Paragenimus westermani, immunogold labeling method was applied using serum immunoglobulins (IgG) of the dog which infected with isolated metacercariae from Cambaroides similis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex(particle size; 12 nm) It was observed by electron microscopy at each tissues of the worm. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were predominantly labeling on the epithelial lamela and lumen of caecum. The above finding showed that antigenic materials in young adult worm tissue were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.365-376
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• 1995

Antigenic localization in Parofonimn iloktsuenensis worm tissues (tegument, intestine and vitelline gland) in different developmental stages of 2 weeks, 3 weeks, 4 weeks, 5 weeks and 33 weeks from albino rats (Sprague-Dawley) infected with P iloktsuenensis was observed by electron microscopy. These worm tissues of different developmental stage of R iloktsuenensis was observed on electromicrograph by immunogold labeling method using R iLoktsuenensis infected rat serum of 10 weeks. Antigenic localization was demonstrated as labeling of gold particles in tissues on electronmicrograph. In tegument, gold particles were labeled on tegumental tissue, generally more numerous on secretory granules in tegumental syncytium 2 weeks than those on the other elder developmental stages, but there was a little variation in antigenicity according to individual worm tissue. In general, antigenicity in tegumental tissue was not strong (gold particles: 0.1-5/1 Mm2). In intestine, a large number of gold particles (15-18/1 Mm2) were labeled in intestinal epithelium. Gold particles were concentrated especially on secretory granules in cytoplasm, and gold particles were labeled not only in cytoplasmic protrusions, but also in intestinal luminal contents. Intencity of labeling of gold particles was not correlated with developmental stage of worms. In vitelline gland, a large number of gold particles were labeled on vitelline globules. The gold particles in vitelline globules (8- 11/1 Mm2) were concentrated in protoplasm among segmental globules . Key words: Pnragonimus iloktsuenensis, immunogold labeling method, tissue antigen ultrastructure.

• 한국전자현미경학회:학술대회논문집
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• 2000.05a
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• pp.23-24
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• 2000

• Journal of Plant Biology
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• v.37 no.4
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• pp.435-439
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• 1994

채종 후 성숙한 인삼 종자 배유에서 cellulase의 국재를 rabbit anti-cellulase와 protein A-gold를 사용한 면역세포화학적 방법을 이용하여 확인하였다. Cellulase의 immunogold particle이 전자밀도가 높은 단백질체와 배유세포벽 주변부에 나타났다. 또한 금입자가 세포벽에 균일하게 분포하였고, 제형층에 접한 분해과정 중의 배유세포벽을 따라 나타났다. 그러나 세포벽의 섬유성 물질과 배유세포의 분해물질로 구성된 제형층에서는 금입자가 관찰되지 않았다.

• Animal cells and systems
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• v.9 no.2
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• pp.65-73
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• 2005

In this study, we found that sperm ball of Urechis unicinctus consisted of a somatic cell and spermatogenic cells. After separation from the sperm ball, individual spermatid floated freely in the coelomic fluid and differentiated into a mature sperm. Because of many nuclear vacuoles, spermatid nucleus was observed to be heterogeneous. Later, the spermatid nucleus condensed into the homogeneous round nucleus of the mature sperm. Perinuclear microtubules could be seen but did not seem to be organized into manchette microtubules. To understand the nature of nuclear condensation during spermiogenesis, the sperm and spermatids (spermiogenic cells) were treated with FITC-phalloidin, or anti-actin-FITC, or labeled with antiactin immunogold particles (AAIP; 10 nm) followed by transmission electron microscopy or confocal laser scanning microscopy. The anti-actin-FITC and FITC-phalloidin reactions occurred distinctly in the nuclei of both spermiogenic cells. FITC-phalloidin reacted more intensely with acrosomes. The AAIP were incorporated mainly into nuclei of both cells sometimes showing local distribution in the nucleus. Nuclear vacuoles of spermatids disappeared progressively with condensation of the nucleus, as the number of incorporated $AAIP/{\mu}m^2$ increased. These results suggest that nuclear actin microfilaments might be closely related to nuclear condensation.

• Korean Journal of Veterinary Service
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• v.20 no.4
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• pp.331-338
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• 1997

The purpose of this study was to establish early diagnosis for bovine virus diarrhea-mucosal disease(BVD-MD) Two Holstein among 22 breeding cows were shown ulcer in the mouth and watery diarrhea. Diarrheal feces and ulcerous lesion of the mouth from 2 cows were sampled for detection of viral antigen. BVD virus was isolated by inoculation of the samples to MDBK cells, and the cytopathic effects were observed in cultured MDBK cells which inoculated with virus isolates from the feces. Viral antigens were detected in the feces and ulceruous lesion by immunogold staining. The serum neutralization titers were shown 1 : 64 or greater in 8 blood samples by using BVD virus (NADL strain). By the RT-PCR, using reverse primer 5'-ACTCCATGTGCCATGTACAG-3', forward primer 5'-ACTCCATGTGCCATGTACAG-3', 285 base pair band specific to BVD virus was detected. In conclusions, the results of above tests which executed using the diarrheal feces and ulcerous lesion of the mouth and the isolates were conformed as BVD virus.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.187-191
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• 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.

• Journal of the Korean Wood Science and Technology
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• v.24 no.1
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• pp.68-74
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• 1996

Present work was undertaken to investigate the origin of milled wood lignin(MWL) in the wood cell wall using immunocytochemical techniques, which can provide the information on the localization of specific antigens(MWL in the present study) to be examined. Spruce MWL dissolved in DMSO and emulsified with Freund adjuvant was injected directly into the mouse spleen. The animals were boostered at two-week intervals after the initial immunization. Blood samples were purified in standard procedures. The characteristics of antibodies against MWL were tested by indirect ELISA. Visualization of MWL was carried out using conventional indirect immunogold-labelling methods on the ultrathin sections of spruce wood. Immuno-TEM observations showed that the immunogold probes were selectively attached to secondary cell walls of spruce wood. The most intense labelling was frequently observed in the S2 layer. In contrast, gold labelling in the lignin-rich regions, such as middle lamella and cell corner was not found. The immuno-TEM provides an indication that spruce MWL originates from the S2 layer.

• Parasites, Hosts and Diseases
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• v.28 no.1
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• pp.11-24
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• 1990

In order to observe the antigenic localization in the tissues of the adult Clonorchis sinensis, immunogold labeling method was applied using serum immunoglobulins (IgG) of either worm·infected rabbits (group I) or antigen-immunized rabbits (group II) (by the body quid obtained from the adult worms). The electron micrographs of the sectioned worm tissue antigens, embedded in Lowicryl HM 20 medium and stained with protein A-gold complex (particle sixte: 12 nm), were compared between the group I and group II. The gold particles were observed in the interstitial matrix of the worm parenchyma, the epithelial lamellae of the cecum, and the cecal lumen both in group I and II. But the particles were in general more concentrated in group II. The gold particles were not observed on the basal lamina of the tegument or on vitelline glands in group I, while they were highly concentrated on those areas in group II. There were also differences in the antigenicity of interstitial matrix(reacted with group I IgG) and head part(reacted with group II IgG) of the sperm cells in the seminal receptacle. Conclusively, it is suggested that the substances comprising the basal lamina of the tegument or vitelline glands act as specific antigens reacting with antigen(body quid) immunized rabbit IgG. On the other hand, the substances in the cecal lumen and cecal epithelial lamellae are thought to be the specific antigen that react with the worm-infected rabbit IgG.