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Messenger RNA Detection in Leukemia Cell lines by Novel Metal-Tagged in situ Hybridization using Inductively Coupled Plasma Mass Spectrometry

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Author(s): Olga I. Ornatsky | Vladimir I. Baranov | Dmitry R. Bandura | Scott D. Tanner | John Dick

Journal: Translational Oncogenomics
ISSN 1177-2727

Volume: 1;
Start page: 1;
Date: 2006;
Original page

Keywords: in situ hybridization | oligonucleotide probes | leukemia cell lines | mass spectrometry | metal-tagged affinity reagents | multiplex gene analysis

ABSTRACT
Conventional gene expression profiling relies on using fluorescent detection of hybridized probes. Physical characteristics of fluorophores impose limitations on achieving a highly multiplex gene analysis of single cells. Our work demonstrates the feasibility of using metal-tagged in situ hybridization for mRNA detection by inductively coupled plasma mass spectrometry (ICP-MS). ICP-MS as an analytical detector has a number of unique and relevant properties: 1) metals and their stable isotopes generate non-overlapping distinct signals that can be detected simultaneously; 2) these signals can be measured over a wide dynamic range; 3) ICP-MS is quantitative and very sensitive. We used commercial antibodies conjugated to europium (Eu) and gold together with biotinylated oligonucleotide probes reacted with terbium-labeled streptavidin to demonstrate simultaneous mRNA and protein detection by ICP-MS in leukemia cells.
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