Fluorescence coding techniques for RNA detection

Fluorescence imaging along with a wide range of fluorescence detection methodologies enables the visualization of macromolecular dynamics in clinical diagnostics and fundamental biomedical research. However, multiplexed fluorescence-based detection has hitherto been largely limited by the overlap of the emission spectra of fluorophores, which only with four or five fluorochrome dyes enable simultaneous use. Fluorescent barcodes with high resolution and high throughput have emerged as a versatile biosensor platform for multianalyte analysis via fine control of fluorescence proportional types, spatial geometries, specific fluorescence properties, and time dimensions. Fluorescence imaging of RNA enables the visualization of physiological and pathological processes at the subcellular level, which is highly important for biomedical diagnostics and therapies and has made tremendous progress toward understanding the complexity of biological systems. In this review, we provide the design and development of fluorescence coding techniques for multiplexed bioanalysis detection and then discuss their applications in RNA biosensors. Additionally, associated challenges in the field and potential solutions will be discussed. Overall, we hope this review will inspire researchers and pave the way for the future design of fluorescent biosensors.