Nucleic Acid Based Fluorescent Nanothermometers

Accurate thermometry at micro- and nanoscales is essential in many nanobiotechnological applications. The nanothermometers introduced in this paper are composed of labeled molecular beacons (MBs) comprising gold nanoparticles (AuNPs) on which, depending on application, many MBs of one or more types are immobilized. In this design, three differently labeled MBs with different thermostabilities function as the sensing elements, and AuNPs act as carriers of the MBs and also quenchers of their fluorophores. This flexible design results in a number of nanothermometers with various temperature-sensing ranges. At the lowest temperature, the MBs are in the closed form, where they are quenched. By increasing the temperature, the MBs start to open with respect to their melting points (Tm), and as a result, the fluorescence emission will increase. The temperature resolution of the nanoprobes over a range of 15-60 degrees C is less than 0.50 degrees C, which indicates their high sensitivity. Such a good temperature resolution is a result of the specific design of the unusual less stable MBs and also presence of many MBs on AuNPs. The reproducibility and precision of the probes are also satisfactory. The multiplex MB nanoprobe is suitable for thermal imaging by fluorescence microscopy.