• Title/Summary/Keyword: Fluorescence in situ hybridization analysis

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국산 Fluorescence in Situ Hybridization 시스템을 이용한 다양한 검체에서의 염색체 분석 (Chromosome Analysis in Clinical Samples by Chromosome Diagnostic System Using Fluorescence in Situ Hybridization)

  • 문신용;방명걸;오선경;류범용;황도영;정병준;최진;손철;장준근;김종원;김석현;최영민
    • Clinical and Experimental Reproductive Medicine
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    • 제24권3호
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    • pp.335-340
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    • 1997
  • Fluorescence in situ hybridization (FISH) techniques allow the enumeration of chromosome abnormalities and from a great potential for many clinical applications. In order to produce quantitative and reproducible results, expensive tools such as a cooled CCD camera and a computer software are required. We have developed a Chromosome Image Processing System (Chips) using FISH that allows the detection and mapping of the genetic aberrations. The aim of our study, therefore, is to evaluate the capabilities of our original system using a black-and-white video camera. As a model system, three repetitive DNA probes (D18Z1, DXZ1, and DYZ3) were hybridized to variety different clinical samples such as human metaphase spreads and interphase nuclei obtained from uncultured peripheral blood lymphocytes, uncultured amniocytes, and germ cells. The visualization of the FISH signals was performed using our system for image acquisition and pseudocoloring. FISH images were obtained by combining images from each of probes and DAPI counterstain captured separately. Using our original system, the aberrations of single or multiple chromosomes in a single hybridization experiment using chromosomes and interphase nuclei from a variety of cell types, including lymphocytes, amniocytes, sperm, and biopsied blastomeres, were enabled to evaluate. There were no differences in the image quality in accordance with FISH method, fluorochrome types, or different clinical samples. Always bright signals were detected using our system. Our system also yielded constant results. Our Chips would permit a level of performance of FISH analysis on metaphase chromosomes and interphase nuclei with unparalleled capabilities. Thus, it would be useful for clinical purposes.

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암모니아 부분산화 공정의 제어와 미생물 군집의 변화 (Influence of FA and FNA to Microbial Community in Sequencing Batch Ammonium Partial Nitrification System)

  • 안조환
    • 한국물환경학회지
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    • 제31권4호
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    • pp.399-406
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    • 2015
  • A sequencing batch reactor was operated under different pH conditions to see the influence of free ammonia (FA) and free nitrous acid (FNA) to microbial community on ammonium partial nitrification. Long-term influences of FA and FNA were evaluated by polymerase chain reaction-denaturing gradient gel electrophoresis and fluorescence in situ hybridization. Nitrite accumulation was successfully achieved at pH 8.2 and 6.3. The shifts in the microbial community were observed when influent ammonia concentration increased to 1 g $NH_4$-N/L at pH 8.2, and then when pH was dropped to 6.3. Both Nitrosomonas and Nitrosospira were selected during the startup of the reactor, and eventually became dominant members as ammonia-oxidizing bacteria. The results of molecular microbiological analysis strongly suggested that the composition of microbial community was changed according to the method used to control nitrite-oxidizing bacteria.

혐기성 SBR을 이용한 anammox 미생물 배양 및 fluorescence in situ hybridization (FISH)을 통 미생물 군집 분석

  • 한동우;윤호준;김동진
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.286-289
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    • 2001
  • Anaerobic ammonium oxidation with nitrite to $N_2$(anammox) is a recently discovered microbial reaction with interesting potential for nitrogen removal from wastewater. Here we investigated the microbial community structure in the sequencing batch reactor(SBR) with an anammox activity. The SBR was optimized for the enrichment of a very slowly growing microbial community and showed that possibility of anaerobic ammonium oxidation. Fluorescence in situ hybridization(FISH) analysis revealed that anaerobic ammonium oxidizers were Candidatus Brocadia anammoxidans and Candidatus Kuenenia stuttgartiensis. Furthermore, Nitrosomol1as spp. of the ${\beta}$ -subclass of Proteobacteria was also present within the anaerobic SBR microorganisms.

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Fluorescence in situ hybridization(FISH) 기법을 이용한 인간 생식세포 및 착상전 배아의 유전이상 검색 (Detection of genetic abnormalities in human sperm, oocytes, and preimplantation embryos using fluorescence in situ hybridization (FISH))

  • 방명걸
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 1998년도 제4차 학술발표대회 및 정기총회
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    • pp.12-18
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    • 1998
  • Tremendous progress has been made over the past quarter-century studying the genetics of gametogenesis and the resulting gametes and embryos. Studies merging molecular techniques and conventional cytogenetics are now beginning to bridge the gap between what we have learned about the meiotic process in males and females and what we know of the mitotic chromosomes of zygotes. Numerical abnormalities in sperm, oocytes and embryo can now diagnosed by fluorescence in situ hybridization (FISH). "At risk" couples can, therefore, have only unaffected embryos replaced in the sterus and avoid the possibility of terminating a pregnancy that might only be diagnosed as affected later gestation. Single-cell genetic analysis has also provided powerful tools for studying genetic defects arising during early human development. Recent studies of sperms, oocytes and cleavage-stage human embryos have revealed an unexpectedly high incidence. These genetic abnormalities are likely to contribute to early pregnancy loss and have important implications for improving pregnancy rates in infertile couples by assisted reproduction. The widespread use of preimplantation genetic diagnosis (PGD) awaits further documentatio of safety and accuracy. Other issues also must be addressed. First, the ethical issues regarding germ cell and embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for non-genetic disorders such as gender selection. Finally, the experimental nature of these procedure must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved assisted reproductive technology success rates may make PGD a more widely use clinical tool. The future awaits these development.velopment.

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3중 DNA probe를 이용한 FISH(fluorescence in situ hybridization) 기법으로 방사선에 의한 염색체 이상 분석 (Analysis of Chromosome aberrations by fluorescence in situ hybridization using triple chromosome-specific probes in human lymphocyte exposed to radiation)

  • 정해원;김수영;하성환
    • Journal of Radiation Protection and Research
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    • 제24권1호
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    • pp.45-53
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    • 1999
  • 각 염색체에 특이한 DNA probe를 이용하는 FISH기법은 방사선에 의해 유발된 상호전좌 및 삽입 등의 염색체의 구조적 변화를 측정하는 매우 효과적인 방법으로서 그 활용성이 증가되고 있다. 본연구는 방사선 피폭시 생물학적 선량측정법으로서 FISH기법을 활용하기 위하여 사람의 1, 2, 4번 염색체에 특이한 probe를 이용하여 고선량 단일 피폭시 유발된 각종 염색체 이상빈도를 관찰하고 이를 PAINT분류체계에 의해 분석하였다. 방사선 조사에 의한 염색체 이상빈도는 상호전좌(t)와 이동원염색체(dic)의 수가 선량 증가에 따라 같이 증가하는 것을 알 수 있으며 color junction의 수도 선량에 따라 증가하는 것을 알 수 있었다. 상호전좌의 빈도는 이동원 염색체의 빈도보다 상대적으로 높게 나타났다. 삽입(ins), 무동원염색체(ace), 및 환상염색체(r)의 수도 선량 증가에 따라 같이 증가하는 것을 알 수 있었다. 기존의 염색체재배열 분석방법과 비교해 볼 때 FISH기법은 다양한 형태의 염색체재해열을 보다 쉽게 관찰할 수 있게 하며 생물학적 선량제로서 중요한 역할을 할 것이라 기대된다.

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두경부 선양낭성암종에서 형광동소결합을 이용한 제17번 염색체의 다염색체 소견 (Fluorescence In Situ Hybridization Analysis for Polysomy of Chromosome 17 in Head and Neck Adenoid Cystic Carcinomas)

  • 최건;박재형;최충식;송재준;정광윤;최종욱
    • 대한두경부종양학회지
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    • 제16권1호
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    • pp.3-8
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    • 2000
  • Background and Objectives: Adenoid cystic carcinoma of salivary glands is characterized by insidious growth over many years, local recurrences, and distant metastasis and classified to three distinct histologic subtypes: tubular, cribriform, and solid. The solid type is known to have the worst prognosis. However, histopathologic heterogeneity is observed in tumors from the same patient. We have attempted to elucidate the genotypic differences, characterized by polysomies of chromosome 17, in adenoid cystic carcinoma according to the phenotypic histopathologic heterogeneity. Materials and Methods: Fluorescence in situ hybridization was performed on formalin-fixed paraffin blocks from seven patients with head and neck adenoid cystic carcinoma, using the centromeric $\alpha$-satellite probe of chromosome 17 to detect nuclei exhibiting polysomy. The difference in polysomeric chromosome expression in cribriform, tubular, solid type and type I, II, III according to the Szanto classification was analyzed. Results: Polysomy of chromosome 17 was found in 15.28% of the cribriform type, in 15.68% of the tubular type, and in 18.87% of the solid type. The proportion of polysomy was statistically higher in the solid type than in the cribriform type(p<0.05), and the proportion of polysomy increased progressively from type 1 to type 3, but this trend was statistically insignificant(p>0.05). Conclusion: We suggest that there may be genetic variations in tumor from the same patient depending on the histopathologic heterogenetiy in adenoid cystic carcinomas.

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GISH와 FISH를 이용한 양파와 파간 종간교잡계통의 염색체 분석 (Chromosome Analysis Using GISH and FISH of Interspecific Hybrids between Allium cepa L. and A. fistulosum L.)

  • 김철우;이을태;김화영;최인후;방진기;구달회;방재욱
    • 한국육종학회지
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    • 제41권4호
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    • pp.468-473
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    • 2009
  • 본 연구에서는 양파와 파간 종간교잡 F1의 불임성의 한 요인으로 여겨지는 염색체 이상과 교잡식물체의 염색체의 조성을 밝혀 향후 종간교잡을 이용한 양파의 형질개량과정에 종간교잡 육종의 효율성을 높이고자 하였다. 종간교잡식물체의 감수분열 염색체를 관찰한 결과 중기에 7쌍의 2가염색체와 2개의 1가염색체가 관찰되었다. 체세포 염색체수는 16개로 양파, 파의 염색체 수와 같았으며 GISH한 결과 파와 양파의 염색체가 각각 8개씩 확인되었다. 5S, 45S, TSD DNA를 probe로 하여 양파, 파, 종간잡종의 염색체의 signal을 관찰한 결과 5S와 45S rDNA는 는 3종 모두에서 1쌍의 염색체에서 나타났는데 5S의 경우 염색체의 subcentromeric 에서 45S는 부수체 염색체에서 관찰되었다. TSD DNA는 염색체의 telomeric 부위에서 관찰되었으나 양파의 경우 2쌍은 염색체의 말단부와 중심체 부분에서 signal이 관찰되었고 종간교잡 세포에서도 2쌍이 관찰되어 양파에서 유래된 염색체임이 확인되었다.

Molecular Identification of the Toxic Alexandrium tamiyavanichii (Dinophyceae) by the Whole-cell FISH Method

  • Kim Choong-Jae;Yoshimatsu Sada-Akfi;Sako Yoshihiko;Kim Chang-Hoon
    • Fisheries and Aquatic Sciences
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    • 제7권4호
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    • pp.175-183
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    • 2004
  • The dinoflagellate Alexandrium tamiyavanichii Balech, a producer of toxins causing paralytic shellfish poisoning (PSP), has recently been considered as one of main organisms responsible for toxication of shellfish in Japan. In this study, A. tamiyavanichii was subjected to a molecular phylogenetic analysis inferred from 28S rDNA D1-D2 sequences and a species-specific LSU rRNA-targeted oligonucleotide DNA probe was designed to identify A. tamiyavanichii using the whole cell-FISH (fluorescence in situ hybridization). The sequences of the 28S rDNA D1-D2 region of A. tamiyavanichii showed no difference from A. cohorticular AF1746l4 (present name A. tamiyavanichii) and formed a distinct clade from the 'tamarensis species complex'. The probe, TAMID2, reacted specifically with A. tamiyavanichii cultured cells, without any cross-reaction with other species belonging to the same genus, including A. tamarense, A. catenella, A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax. In a test of cross-reactivity with a field sample, TAMID2 reacted consistently with only A. tamiyavanichii, indicating that the present protocol involving the TAMID2 probe might be useful for detecting toxic A. tamiyavanichii in a simple and rapid manner.