Nucleic acid biomarkers, which involve in gene expression control, are found specific for many kinds of cancers. Förster Resonance Energy Transfer (FRET) based applications are one of the most promising for nucleic acid biosensing. As parallel detection of multiple nucleic acids is highly demanded and spectral multiplexing is limited by optical crosstalk, temporal multiplexing is used for opening another dimension of the multiplexing. We focus on developing different Tb-to-dye FRET distances to create specific intensity signals corresponding to different nucleic acid sequences. The Tb-dye distances can be tuned by the specific location of the Tb donor using different lengths of DNA. Amplification technologies, such as hybridization chain reaction (HCR) and rolling circle amplification (RCA), are used to achieve simplicity, rapidity, selectivity, and sensitivity of nucleic acid detection. Temporal multiplexing FRET was also combined with spectral (color) multiplexing for higher-order multiplexed detection. Moreover, a single Tb-QD FRET modeling demonstrated the possibility of nanoparticle-based temporal multiplexing.

Time-resolved multiplexed förster resonance energy transfer for nucleic acid biosensing

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ACS Fall 2022

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