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  • 1
    Electronic Resource
    Electronic Resource
    Detection of Genetic Aberrations in Bladder Cancer Using in Situ Hybridization (1993)
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 677 (1993), S. 0 
    Details
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1749-6632.1993.tb38778.x
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  • 2
    Electronic Resource
    Electronic Resource
    Improved mRNA in situ hybridization on formaldehyde-fixed and paraffin-embedded tissue using signal amplification with different haptenized tyramides (1998)
    Springer
    Histochemistry and cell biology 110 (1998), S. 571-577 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We report an optimized in situ hybridization (ISH) protocol with a rapid signal amplification procedure based on catalyzed reporter deposition (CARD) to increase the sensitivity of non-isotopic mRNA ISH on formaldehyde-fixed and paraffin-embedded tissue. The CARD method is based on the deposition of haptenized tyramide molecules in the vicinity of hybridized probes catalyzed by horseradish peroxidase. Commercially available and newly synthesized haptenized tyramides, including digoxigenin-, biotin-, di- and trinitrophenyl- as well as fluorescein-tyramide, were compared. The haptenized tyramides were visualized using peroxidase conjugated anti-hapten antibodies followed by the diaminobenzidine reaction. As a test system, we applied digoxigenin-labeled oligonucleotides to detect insulin and vasoactive intestinal polypeptide mRNA in pancreatic endocrine tumors and liver metastases. Our results indicate that specificity, sensitivity, and applicability of oligonucleotide mRNA ISH can be significantly improved by using chemically digoxigenin-labeled oligonucleotide probes and signal amplification by CARD. Furthermore, all tested tyramides provided approximately equal amplification efficiency. In conclusion, CARD signal amplification should further promote mRNA ISH studies on paraffin-embedded tissues and allow for multiple-target nucleic acid detection in situ.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004180050319
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  • 3
    Electronic Resource
    Electronic Resource
    Multicolour preparations for in situ hybridization using precipitating enzyme cytochemistry in combination with reflection contrast microscopy (1993)
    Springer
    Histochemistry and cell biology 100 (1993), S. 357-366 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have further developed a method for the detection of different enzyme cytochemical reaction products by means of reflection contrast microscopy (RCM). By embedding these enzyme precipitates in a protein matrix, we were able to prevent the reaction products from dissolving in immersion oil, which is required for RCM analysis. The applicability of the RCM procedure is, therefore, extended to a range of cytochemical enzyme precipitation methods, which normally result in oil soluble reaction products. To test their usefulness, these enzyme precipitates have been used in single- as well as double-label in situ hybridization (ISH) procedures to visualize a number of DNA target sequences by several different reflection colours, i.e. white, yellow and red. Three repetitive DNA probes for the (sub)centromeric regions of chromosomes 1, 7 and 17, as well as a repetitive DNA probe for the telomeric region of chromosome 1, and two cosmid DNA probes (40 kb each) for both arms of chromosome 11 could be detected with high efficiency in both interphase and metaphase preparations. Moreover the enzyme precipitates were shown to be stable upon exposure to excitation light or upon storage. It may be concluded that these findings render RCM a sensitive method for the visualization of multiple targets in biological specimens.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00268934
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  • 4
    Electronic Resource
    Electronic Resource
    Fluorescence in situ hybridization on paraffin-embedded abortion material as a means of retrospective chromosome analysis (1994)
    Springer
    Human genetics 〈Berlin〉 94 (1994), S. 518-522 
    Details
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A fluorescence in situ hybridization (FISH) procedure was used to detect chromosome abnormalities in archival abortion material. Nuclei were isolated from 50-μm-thick tissue blocks from 18 selected and karyotyped abortions. Five probes for repetitive centromeric sequences of chromosomes 1, 16, 18, X and Y were used. For each chromosome, at least 200 nuclei were scored blindly, i.e. without knowledge of the karyotype. The FISH results obtained were compatible with the cytogenetic data in 14 cases. There were four discrepancies. Two of these were observed for cases karyotyped as trisomy 16. Furthermore, FISH results showed trisomy 18 in two cases having normal chromosomes 18 and 18q+, respectively. The latter case was not discrepant if the structural rearrangement involved chromosome 18 material. The remaining discrepancies could be explained by chromosomal mosaicism. Admixture of normal maternal cells was also noted. It is concluded that FISH can be used to study retrospectively the presence of chromosome abnormalities in abortion material. However, the quality obtained after the use of fresh material is superior.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00211018
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  • 5
    Electronic Resource
    Electronic Resource
    Presence of chromosomal mosaicism in abnormal preimplantation embryos detected by fluorescence in situ hybridisation (1994)
    Springer
    Human genetics 〈Berlin〉 94 (1994), S. 609-615 
    Details
    ISSN: 1432-1203
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The extent of chromosomal mosaicism in human preimplantation embryos was examined using an improved procedure for the preparation and spreading of interphase nuclei for use in fluorescence in situ hybridisation, allowing the analysis of every nucleus within an embryo. One cell showed no hybridisation signals in only three of the 38 embryos that were included in this study, i.e. the hybridisation efficiency per successfully spread nucleus was 99% (197/200). Double-target in situ hybridisation analyses with X- and Y-chromosome-specific probes was performed to analyse nine embryos resulting from normal fertilisation, 22 polypronucleate embryos and seven cleavage-stage embryos where no (apronucleate) or only one pronucleus (monopronucleate) was observed. We also analysed autosomes 1 and 7 by double-target in situ hybridisation in the nuclei of two apronucleate, one monopronucleate and four polypronucleate embryos. All nine embryos that resulted from normal fertilisation were uniformly XY or XX. None of the apronucleate or monopronucleate embryos was haploid: three were diploid, one was triploid and three were mosaic. Fertilisation was detected by the presence of a Y-specific signal in four of these embryos. Of the polypronucleate embryos, two were diploid, two were triploid and 18 were mosaic for the sex chromosomes and/or autosomes 1 and 7. These results demonstrate that fertilisation sometimes occurs in monopronucleate embryos and that chromosomal mosaicism can be detected with high efficiency in apronucleate, monopronucleate and polypronucleate human embryos using fluorescence in situ hybridisation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00206952
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  • 6
    Electronic Resource
    Electronic Resource
    Detection of genomic changes in cancer byin situ hybridization (1994)
    Springer
    Molecular biology reports 19 (1994), S. 31-44 
    Details
    ISSN: 1573-4978
    Keywords: interphase cytogenetics ; chromosome aberrations ; cancer ; DNA probes ; comparative genomic hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00987320
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  • 7
    Electronic Resource
    Electronic Resource
    Cytochemical detection systems for in situ hybridization, and the combination with immunocytochemistry. ‘Who is still afraid of red, green and blue?’ (1995)
    Springer
    Journal of molecular histology 27 (1995), S. 833-858 
    Details
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An overview is given of the different non-radioactive cytochemical detection methodologies that are currently utilized to localize nucleic acid sequences in chromosomes, cells and tissue sections. Dependent on the reporter molecule (fluorochrome, enzyme or hapten) that is used to modify the appropriate nucleic acid probe, and the sensitivity that is required, the in situ hybridized sequences can be detected either directly after hybridization or indirectly, using cytochemical detection and amplification layers. These may then contain antibody and/or avidin molecules conjugated to fluorochromes, enzymes or colloidial gold particles. Since the choice of a suitable probe-labelling method in combination with a fluorescence, enzyme cytochemical or immunogold-silver detection procedure is often determined by the user's own practical experience and applications, the different detection methodologies are compared with each other in detail with respect to sensitivity, resolution, applicability for multiple probe detection, and signal evaluation. Furthermore, procedures are reviewed for the combination of in situ hybridization with immunocytochemical detection of proteins and/or incorporated bromodeoxyuridine, which allow the simultaneous visualization of genomic phenotypic and/or cell cycle parameters in the same sample. Possible improvements with respect to sensitivity, specificity and multiplicity of the detection methods, which may be interesting for one's own experimental design, are finally being discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00173839
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  • 8
    Electronic Resource
    Electronic Resource
    Cytochemical detection systems forin situ hybridization, and the combination with immunocytochemistry. ‘Who is still afraid of red, green and blue?’ (1995)
    Springer
    Journal of molecular histology 27 (1995), S. 833-858 
    Details
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An overview is given of the different non-radioactive cytochemical detection methodologies that are currently utilized to localize nucleic acid sequences in chromosomes, cells and tissue sections. Dependent on the reporter molecule (fluorochrome, enzyme or hapten) that is used to modify the appropriate nucleic acid probe, and the sensitivity that is required, thein situ hybridized sequences can be detected either directly after hybridization or indirectly, using cytochemical detection and amplification layers. These may then contain antibody and/or avidin molecules conjugated to fluorochromes, enzymes or colloidial gold particles. Since the choice of a suitable probe-labelling method in combination with a fluorescence, enzyme cytochemical or immunogold-silver detection procedure is often determined by the user’s own practical experience and applications, the different detection methodologies are compared with each other in detail with respect to sensitivity, resolution, applicability for multiple probe detection, and signal evaluation. Furthermore, procedures are reviewed for the combination ofin situ hybridization with immunocytochemical detection of proteins and/or incorporated bromodeoxyuridine, which allow the simultaneous visualization of genomic phenotypic and/or cell cycle parameters in the same sample. Possible improvements with respect to sensitivity, specificity and multiplicity of the detection methods, which may be interesting for one’s own experimental design, are finally being discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02389591
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