Nanoparticles as probes for imaging mass-cytometry analysis of microRNA

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The core technology of Fluidigm Canada Inc. is mass cytometry that enables real-time quantitative analysis ofbiomarkers in single cells. The technology is premised on the use of stable isotopes as labels whereby thetagged cells are introduced into an inductively coupled plasma and the ion cloud is analyzed by time-of-flightmass spectrometry. Amplification methods based on a cascaded hybridization assay are employed for analyzingmessenger RNA and proteins; however, this approach is ineffective for short nucleic acid targets such asmicroRNA. Hence alternative signal amplification strategies with high specificity and high detection signalsare of interest. The proposed work investigates DNA-functionalized nanoparticles that will cross-link in vitroin the presence of microRNA as probes for mass cytometry. The cellular uptake, cross-linking and subsequentexcretion or removal of the nanoparticles will be investigated. With each nanoparticle consisting of thousandsof atoms, variations in the number of nanoparticles in cells as a result of differences in the extent ofaggregation will give large differences in the signal. The correlation between microRNA level and masscytometry signal will be examined. The project will expand the applicability of mass cytometry beyondproteomics and messenger RNA and further empower biomedical research that investigates the role of miRNAin various diseases, including cancer, pre-eclampsia and cardio vascular diseases.