MicroRNA-21 expression in single living cells revealed by fluorescence and SERS dual-response microfluidic droplet platform

Abstract

Analysis of single-cell microRNA is essential to reveal cell heterogeneity at the genetic level. It raises a high demand for single-cell analytical methods because single-cell microRNA sequences are highly similar and small in size and feature low-level expression. Herein, SERS and fluorescence imaging technology were introduced into a microfluidic droplet platform to realize direct in situ, nondestructive, and highly sensitive detection of a small number of microRNA-21 (miR-21) in a single intact living cell. A multifunctional plasmonic nanoprobe was designed by decorating a gold nanoparticle with fluorescent dye (ROX)-labeled probe DNA and capture DNA strands. The dual-signal switching of fluorescence turn-off and SERS turn-on of ROX in response to miR-21 achieves highly sensitive and reliable detection of miR-21 in a single cell. The turn-on of SERS signal with a zero background guarantees the sensitivity of the detection. The fluorescence-SERS simultaneous response strategy was able to mutually corroborate the test results, improving the reliability of determining low-level expression of miR-21. SERS combined with encapsulation of microdroplets provides a feasible way to conduct in situ, nondestructive determination of miR-21 secreted by single cells, avoiding cell lysis and tedious time-consuming steps of miR-21 isolation. As a result, the miR-21 expressed by various types of single cells was investigated by fluorescence imaging and the cellular heterogeneity in miR-21 expression was evaluated accurately and quantitatively by SERS. This research would provide important reference information for understanding the effects of miRNAs on cancer diseases at the single-cell level.