• Title/Summary/Keyword: fluorescence in situ hybridization (FISH)

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Detection of Chromosomal Rearrangements by Chromium in Human Lymphocyte Using Fluorescence in situ Hybridization (FISH) with Triple Combination of Composite whole Chromosome Specific Probe (FISH(fluorescence in situ hybridization)를 이용하여 분석한 크롬에 의해 유발된 염색체 이상)

  • 정해원;김수영;맹승희;이용묵;유일재
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.14-19
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    • 1999
  • Chromosome rearrangements induced in human lymphocyte after in vitro exposure to chromium were analysed by the use of fluorescence in situ hybridization(FISH) with triple combination of composite whole chromosome-specific probe for chromosome 1, 2 and 4. Chromosome aberrations was scored by the Protocol for Aberration Identification and Nomenclature Terminology (PAINT). Stable translocation was the most frequent type of aberrations and dicentrics and insertions were also observed. Chromium treatment enhanced the frequencies of stable translocations and color junctions in a dose-dependent manners, but no distinct increase of dicentrics and insertions was seen. The ratio of the yields of translocation to the yields of dicentric varied between 13 to 27. The presents results demonstrate fluorescent in situ hybridization (FISH) is useful for detecting chromosomal rearrangements induced by chromium.

Chromosome Rearrangements Detected by Fluorescence in situ Hybridization in Human Lymphocyte Exposed to Bleomycin (Fluorescence in situ hybridization (FISH)를 이용하여 분석한 Bleomycin에 의한 사람 림프구의 염색체 재배열)

  • 손은희;정경인;정해원
    • Environmental Mutagens and Carcinogens
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    • v.17 no.1
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    • pp.12-16
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    • 1997
  • Chromosome rearrangement induced by bleomycin were identified by fluorescence in situ hybridization with probe for chromosome 4. The frequency of color junctions, translocations, dicentric and acenttic fragments increased with bleomycin dose. Different types of balanced translocation and dicentric were scored and compared. The frequency of cells exhibiting multiple aberration was higher compared to that of cells exposed to Gamma radiation suggesting that effect of bleomycin might be similar to that of high LET radiation.

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Radiation induced Chromosome aberration in human Iymphocyte detected by Fluorescence in sifu hybridization (FISH(Fluorescence in situ hybridization)를 이용하여 분석한 방사선에 의해 유발된 림프구의 염색체 이상)

  • 정해원;손은희;기혜성;하성환
    • Environmental Mutagens and Carcinogens
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    • v.16 no.2
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    • pp.88-96
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    • 1996
  • Fluorescence in situ hybridization (FISH) with the DNA probe for human chromosome 4 was used to analyse in vitro radiation induced chromosome rearrangement in peripheral lymphocyte. Translocations, dicentrics, acentrics and color junctions involving the painted chromosome were scored according to the Protocol for Aberration Identification and Nomenclature Terminology (PAINT) system. The frequency of chromosome rearrangements including reciprocal translocation, dicentric, acentric fragment and color junction increased with radiation dose. The frequency of dicentric chromosome reduced by the fixation time following irradiation, whereas that of translocation was relatively persistent. The applicability of FISH for scoring stable translocation for biological dosimetry was demonstrated.

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Single-molecule fluorescence in situ hybridization: Quantitative imaging of single RNA molecules

  • Kwon, Sunjong
    • BMB Reports
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    • v.46 no.2
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    • pp.65-72
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    • 2013
  • In situ detection of RNAs is becoming increasingly important for analysis of gene expression within and between intact cells in tissues. International genomics efforts are now cataloging patterns of RNA transcription that play roles in cell function, differentiation, and disease formation, and they are demon-strating the importance of coding and noncoding RNA transcripts in these processes. However, these techniques typically provide ensemble averages of transcription across many cells. In situ hybridization-based analysis methods complement these studies by providing information about how expression levels change between cells within normal and diseased tissues, and they provide information about the localization of transcripts within cells, which is important in understanding mechanisms of gene regulation. Multi-color, single-molecule fluorescence in situ hybridization (smFISH) is particularly useful since it enables analysis of several different transcripts simultaneously. Combining smFISH with immunofluorescent protein detection provides additional information about the association between transcription level, cellular localization, and protein expression in individual cells.

Detection of Benzene Metabolite Induced Aneuploidy and Translocation in HL-60 Cells by Fluorescence in situ Hybridization using Whole Chromosome-specific Probes for Chromosome 8 and 21 (벤젠 대사산물에 의해 유도된 HL-60 세포의 8번 및 21번 염색체의 이수성 및 상호전좌)

  • 김수영;정해원
    • Environmental Mutagens and Carcinogens
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    • v.22 no.2
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    • pp.90-96
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    • 2002
  • Benzene is a widespread human carcinogen, inducing leukemia and hematotoxicity. Exposure to benzene metabolites has been shown to cause genetic damage, including aneusomy and chromosome aberrations. Fluorescence in situ hybridization(FISH) procedure was used to determine if the benzene metabolite, 1, 2, 4-benzenetriol(BT), hydroquinone(HQ) and trans, trans-muconic acid(t,t-MA) induced specific chromosomal change in HL-60 cells. Treatment with BT, HQ and t,t-MA resulted in the induction of monosomy 8 and 21 in HL-60 cells in a dose-dependent manner. All of these metabolites also induced trisomy 8 and 21, but no correlation between frequencies of trisomy and concentration was found. Translocations between chromosome 8 and another unidentified chromosome [t(8:\ulcorner)], and between chromosome 21 and another unidentified chromosome [t(8:21)] were found. However, translocation between chromosome 8 and 21 [t(8:21)] was not found. Results indicate that the benzene metabolites, BT, HQ and t,t-MA, induce chromosome specific numerical and structural aberrations, and the fluorescence in situ hybridization (FISH) approach may be a useful and powerful technique for detection of aneuploidy.

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Sex Determination of In Vitro Fertilized Bovine Embryos by Fluorescence In Situ Hybridization Technique

  • Han, M.S.;Cho, E.J.;Ha, H.B.;Park, H.S.;Sohn, S.H.
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.133-137
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    • 2004
  • Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. To evaluate Y-chromosome specificity of the FISH probe, metaphase spreads of whole embryos and lymphocytes were prepared and tested. A male-specific signal was detected on 100% of Y chromosome bearing metaphase specimens. Using the FISH technique with a bovine Y-specific probe, 232 whole embryos of 8 cell- to blastocyst-stage were analyzed. Observing the presence of the Y-probe signal on blastomeres, 102 embryos were predicted as male, and 130 embryos as female. The determining rate of embryo sex by FISH technique was about 93% regardless of embryonic stages. In conclusion, the FISH using a bovine Y-specific DNA probe is an accurate, reliable and quick method for determining the sex of bovine embryos.

Sex Determination of In Vitro Fertilized Bovine Embryos by Fluorescence in situ Hybridization Technique

  • Han, M. S.;E. J. Cho;H. B. Ha;Park, H. S.;S. H. Sohn
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.287-287
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    • 2004
  • Sexing from bovine embryos which were fertilized in vitro implicate a possibility of production of the sex controlled cattle. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. (omitted)

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Frequency of Chromosome Aberrations Detected by Fluorescence in Situ Hybridization Using Triple Chromosome-Specific Probes in o Healthy Korean Population (3중 염색체 probe를 이용한 FISH(fluorescence in situ hybridization)기법으로 분석한 정상인의 염색체 이상빈도)

  • 정해원;김수영;신은희
    • Environmental Mutagens and Carcinogens
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    • v.18 no.2
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    • pp.109-115
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    • 1998
  • Fluorscence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by chemical and physical agents. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to use the FISH method as a biodosimeter for monitoring human population exposed to various chemical and physical agent, baseline level of chromosome rearragement was established. Blood from forty four healthy adults were collected and analysed with whole chromosome-specific probes by human chromosome 1,2 and 4. The frequencies of stable translocation were 2.45 per 100 cell equivalent and those of insertion, color juction, acentric and dicentric were 0.32, 3.28, 0.23 and 0.27 per 100 cell equivalent respectively. The frequencies of chromosome rearragements increased with age in both sexes except for dicenrics. From above result, stable aberrations accumulate with age and it may reflect integrated lifetime exposure of adverse environment.

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Community structure analysis of nitrifying biofilms by 16S rRNA targeted probe and fluorescence in situ hybridization (FISH)

  • Han, Dong-U;Kim, Dong-Jin
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.282-285
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    • 2001
  • The microbial community structure and in situ spatial distribution of ammonia oxidizing and nitrite oxidizing bacteria in nitrifying biofilm of an upflow biological aerated filter system were investigated. The reactor had been continuously operated under high free ammonia concentration and low DO concentration for nitrite accumulation more than 2 years before the experiment. Fluorescence in situ hybridization

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Age Prediction in the Chickens Using Telomere Quantity by Quantitative Fluorescence In situ Hybridization Technique

  • Kim, Y.J.;Subramani, V.K.;Sohn, S.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.5
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    • pp.603-609
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    • 2011
  • Telomeres are special structures at the ends of eukaryotic chromosomes. Vertebrate telomeres consist of tandem repeats of conserved TTAGGG sequence and associated proteins. Birds are interesting models for molecular studies on aging and cellular senescence because of their slow aging rates and longer life spans for their body size. In this longitudinal study, we explored the possibility of using telomeres as an age-marker to predict age in Single Comb White Leghorn layer chickens. We quantified the relative amount of telomeric DNA in isolated peripheral blood lymphocytes by the Quantitative Fluorescence in situ Hybridization technique on interphase nuclei (IQ FISH) using telomere-specific DNA probes. We found that the amount of telomeric DNA (ATD) reduced significantly with an increase in chronological age of the chicken. Especially, the telomere shortening rates are greatly increased in growing individuals compared to laying and old-aged individuals. Therefore, using the ATD values obtained by IQ FISH we established the possibility of age prediction in chickens based on the telomere theory of aging. By regression analysis of the ATD values at each age interval, we formulated an equation to predict the age of chickens. In conclusion, the telomeric DNA values by IQ FISH analyses can be used as an effective age-marker in predicting the chronological age of chickens. The study has implications in the breeding and population genetics of poultry, especially the reproductive potential.