Multiparameter in situ analysis of trophoblast cells in mixed cell populations by combined DNA and RNA in situ hybridization

We developed a non-radioactive assay for simultaneous detection of cytoplasmic mRNA and nuclear genomic DNA in fetal trophoblast cells by sequential in situ hybridization. Trophoblast-specific mRNA is detected with a digoxigenin-labeled RNA probe complementary to HLA-G, followed by visualization thr...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 1995-07, Vol.43 (7), p.709-714
Main Authors: Van Wijk, IJ, Van Vugt, JM, Konst, AA, Mulders, MA, Nieuwint, AW, Oudejans, CB
Format: Article
Language:English
Subjects:
DNA - analysis
DNA Probes
Histocompatibility Antigens Class I - analysis
HLA Antigens - analysis
HLA-G Antigens
Humans
In Situ Hybridization
Luminescent Measurements
Microscopy, Fluorescence
Prenatal Diagnosis - methods
RNA Probes
RNA, Messenger - analysis
Trophoblasts - chemistry
Trophoblasts - cytology
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Summary:We developed a non-radioactive assay for simultaneous detection of cytoplasmic mRNA and nuclear genomic DNA in fetal trophoblast cells by sequential in situ hybridization. Trophoblast-specific mRNA is detected with a digoxigenin-labeled RNA probe complementary to HLA-G, followed by visualization through the generation of stable contrast-rich DAB/Ni complexes. Genomic target DNA is subsequently visualized in labeled cells by fluorescent in situ hybridization using biotin-labeled chromosome-specific DNA probes. Simultaneous visualization of both targets is made possible using a fluorescence microscope with FITC filter and conventional brightfield light. This method allows detection of trophoblast cells within a mixed cell population and, at the same time, analysis of chromosome anomalies in the trophoblast cells identified. For prenatal diagnosis of fetal cells enriched from maternal peripheral blood during pregnancy, this multiparameter in situ analysis of immobilized fetal trophoblast cells will be very useful.
ISSN:0022-1554
1551-5044
DOI:10.1177/43.7.7608525