Bead-Based Flow Cytometric miRNA Expression Profiling

Abstract

The bead-based flow cytometric miRNA expression profiling method involves coupling of miRNA capture probes to carboxylated 5-micron polystyrene beads, RT-PCR amplification of miRNAs, hybridization of miRNAs to the capture beads, staining with streptavidin-phycoerythrin, and and final read out using a flow cytometer to measure bead color (denoting miRNA identity) and phycoerythrin intensity (denoting miRNA abundance). The technique was invented by Lu et al in Broad Institute of MIT and Harvard (Cambridge, Massachusetts, USA) (Lu et al. 2005). The bead-based method has been applied to conduct a systematic expression analysis of 217 mammalian miRNAs from 334 samples, including multiple human cancers. The data proved it is able to distinguish tumors of different developmental origin, to distinguish tumors from normal tissues, and to distinguish tumors of different stages. The technique is feasible, and has the attractive properties of improved accuracy, high speed and low cost. Moreover, it is an efficient detection platform for high throughput miRNA profiling in a quantitative manner. The bead-based miRNA detection method is also easy to implement in a routine clinical setting.