Abstract
Abstract We describe a novel assay that enables analysis of microRNA, mRNA, and protein expression in single cells by flow cytometry. This in situ hybridization-based method employs pairs of target-specific probes, branched DNA amplification technology, and fluorochrome-conjugated label probes, resulting in sensitive detection of different types of RNA targets including mRNA, microRNA, long non-coding RNA and viral RNA. To broaden our understanding of the applicability of this assay, we determined its limit of detection for mRNA and microRNA. Using in vitro bone marrow-derived macrophages as a model, we demonstrate how this assay can be used to study the process of cell differentiation and maturation. This novel assay enables the simultaneous detection of up to four RNA targets in the presence of fluorochrome-conjugated antibodies in single cells and is a powerful new tool that will enable new insights into cellular processes.
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