Abstract
Most of the techniques for simultaneous visualization of proteic cell components and DNA probes are difficult to carry out. We have developed an alternative protocol for simultaneous visualization of DNA probes and cell surface antigens in human cells. The method exploits microwave energy to denature DNA and to anneal the probe after antigen fluorescent detection of unfixed cells previously embedded in a microgel. CD3 antigens and whole genome DNA probes or specific repetitive DNA sequences were colocalized in peripheral blood samples. The results show a strong, specific and consistent hybridization pattern in each cell that allowed correlation between cell subtype (PROTEINS) and nuclear phenotype (DNA).
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Buño I, Diez-Martín JL, Fernández JL, Gosálvez J (1999) Polymorphisms for the size of heterochromatic regions allow sex-independent quantification of post-bmt chimerism targetting metaphase and interphase cells. Haematologica 84: 138–141.
Durm M, Haar FM, Hausmann M, Ludwig H, Cremer C (1997) Optimized Fast-FISH with alpha-satellite probes: acceleration by microwave activation. Braz J Med Biol Res 1: 15–23.
Fernández JL, Goyanes V, Ramiro-Diez J, Gosálvez J (1998) Application of FISH for in situ detection and quantification of DNA breakage. Cytogenet Cell Genet 82: 251–256.
Kögler G, Wolf HH, Heyl A, Arkesteijn G, Wernet P (1995) Detection of mixed chimerism and leukemic relapse after allogenic bone marrow transplantation in subpopulation of leukocytes by Luorescent in situ hybridization in combination with the simultaneous immunophenotypic analysis of interphase cells. Bone Marrow Trasplant 15: 41–48.
Meulemeester de M, Vink A, Jakobs M et al. (1996) The application of microwave denaturation in comparative genomic hybridization. Genet Anal 5: 129–133.
Speel EJ, Herbergs J, Remaekers FC, Hopman AH (1994) Combined immunocytochemistry and Luorescence in situ hybridization for simultaneous tricolor detection of cell cycle, genomic, and phenotypic parameters of tumor cells.J Histochem Cytochem 7: 961–966.
Teerenhovi L, Knuutila S, Ekblom M et al. (1984) A method for simultaneous study of the karyotype morphology and immunologic phenotype of mitotic cells in hematologic malignancies. Blood 64: 1116–1122.
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Gosálvez, J., De La Torre, J., Pita, M. et al. FISHing in the microwave: the easy way to preserve proteins. I. Colocalization of DNA probes and surface antigens in human leukocytes. Chromosome Res 10, 137–143 (2002). https://doi.org/10.1023/A:1014901218648
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DOI: https://doi.org/10.1023/A:1014901218648