Issue 17, 2015

Fluorescent microRNA biosensors: a comparison of signal generation to quenching

Abstract

Many microRNA biosensor platforms use fluorescence signal generation or quenching; however, signal generation is often regarded as the superior method. An argument can be made that if the noise is the same for both methods, then there should be no difference between the two methods. Current literature details the analytical figures of merit (FOM) for transduction and recognition mechanisms that use either signal generation or quenching, but lacks a direct comparison using the same fluorescent reporter molecule. Here we provide such a direct comparison. The signal-on and signal-off fluorescence metrics were found to be comparable rather than competitive. We found fluorescence enhancement provides marginal improvements to sensitivity and limits of detection (LOD) over fluorescence quenching. In fact, both transduction mechanisms are capable of picomolar LOD. The role thermodynamics plays on the sensitivity and LOD are discussed. Both signal-on and signal-off gave statistically similar signal-to-noise ratios. Finally, the selectivity of the two recognition mechanisms for miRNA detection will be addressed. In the future, we will use this knowledge to advance highly sensitive and selective in situ microRNA sensors for cell and tissue imaging.

Graphical abstract: Fluorescent microRNA biosensors: a comparison of signal generation to quenching

Supplementary files

Article information

Article type
Paper
Submitted
06 جمادى الأولى 1436
Accepted
16 رجب 1436
First published
17 رجب 1436

Anal. Methods, 2015,7, 7296-7310

Author version available

Fluorescent microRNA biosensors: a comparison of signal generation to quenching

C. K. Almlie, N. E. Larkey and S. M. Burrows, Anal. Methods, 2015, 7, 7296 DOI: 10.1039/C5AY00504C

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements