title: Messenger RNA Detection in Leukemia Cell lines by Novel Metal-Tagged in situ Hybridization using Inductively Coupled Plasma Mass Spectrometry.
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.
authors: IBBME, University of Toronto, Toronto, ON, Canada.
authors: Olga I
authors: OI
authors: Ornatsky
authors: Vladimir I
authors: VI
authors: Baranov
authors: Dmitry R
authors: DR
authors: Bandura
authors: Scott D
authors: SD
authors: Tanner
authors: John
authors: J
authors: Dick
keywords: in situ hybridization; leukemia cell lines; mass spectrometry; metal-tagged affinity reagents; multiplex gene analysis; oligonucleotide probes
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