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

I J Van Wijk, J M Van Vugt, A A Könst, M A Mulders, A W Nieuwint, C B Oudejans

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)709-14
Number of pages6
JournalThe Journal of Histochemistry and Cytochemistry
Volume43
Issue number7
Publication statusPublished - Jul 1995

Cite this

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title = "Multiparameter in situ analysis of trophoblast cells in mixed cell populations by combined DNA and RNA in situ hybridization",
abstract = "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.",
keywords = "DNA/analysis, DNA Probes, HLA Antigens/analysis, HLA-G Antigens, Histocompatibility Antigens Class I/analysis, Humans, In Situ Hybridization, Luminescent Measurements, Microscopy, Fluorescence, Prenatal Diagnosis/methods, RNA Probes, RNA, Messenger/analysis, Trophoblasts/chemistry",
author = "{Van Wijk}, {I J} and {Van Vugt}, {J M} and K{\"o}nst, {A A} and Mulders, {M A} and Nieuwint, {A W} and Oudejans, {C B}",
year = "1995",
month = "7",
language = "English",
volume = "43",
pages = "709--14",
journal = "The Journal of Histochemistry and Cytochemistry",
issn = "0022-1554",
publisher = "Histochemical Society Inc.",
number = "7",

}

Multiparameter in situ analysis of trophoblast cells in mixed cell populations by combined DNA and RNA in situ hybridization. / Van Wijk, I J; Van Vugt, J M; Könst, A A; Mulders, M A; Nieuwint, A W; Oudejans, C B.

In: The Journal of Histochemistry and Cytochemistry, Vol. 43, No. 7, 07.1995, p. 709-14.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Multiparameter in situ analysis of trophoblast cells in mixed cell populations by combined DNA and RNA in situ hybridization

AU - Van Wijk, I J

AU - Van Vugt, J M

AU - Könst, A A

AU - Mulders, M A

AU - Nieuwint, A W

AU - Oudejans, C B

PY - 1995/7

Y1 - 1995/7

N2 - 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.

AB - 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.

KW - DNA/analysis

KW - DNA Probes

KW - HLA Antigens/analysis

KW - HLA-G Antigens

KW - Histocompatibility Antigens Class I/analysis

KW - Humans

KW - In Situ Hybridization

KW - Luminescent Measurements

KW - Microscopy, Fluorescence

KW - Prenatal Diagnosis/methods

KW - RNA Probes

KW - RNA, Messenger/analysis

KW - Trophoblasts/chemistry

M3 - Article

VL - 43

SP - 709

EP - 714

JO - The Journal of Histochemistry and Cytochemistry

JF - The Journal of Histochemistry and Cytochemistry

SN - 0022-1554

IS - 7

ER -