Novel Ratiometric Fluorescent Nanothermometers Based on Fluorophores-Labeled Short Single-Stranded DNA

Novel ratiometric fluorescent short single-stranded DNA (ssDNA) nanothermometers (ssDNA FT) were developed using the fluorescence resonance energy transfer (FRET) effect of the ssDNA's end labeled fluorophores. An optimal ssDNA sequence and associated ssDNA FT were determined through combined MD simulation and temperature-related FRET analysis. Their fluorescence properties and thermo-responsivities were analyzed using fluorescence spectra. The influences of ssDNA' sequence length, sequence composition and fluorescent labels for temperature sensing were investigated. Results revealed the prepared, optimized ssDNA FT showed a high average temperature sensitivity of 7.04% °C^1-, wide linear response range of 0-100 °C, and excellent stability with various environmental factors. Furthermore, this ssDNA FT was successfully used for intracellular temperature sensing in cancer cells and was used for in vivo thermos-imaging during microwave hyperthermia of tumor tissue. Advantages in size, sensitivity, and stability proved the feasibility of ssDNA FT in nanoscale thermometry applications, and this novel fluorescent thermometry mechanism is of large potential in the development of FTs. This investigation of ssDNA's molecular thermosensitivity could give rise to a new prospective in the nanothermometry field.