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By developing lithium-ion-active aerolysin, for the first time we have achieved aerolysin detection of single-stranded DNA longer than 100 nt., Wild-type aerolysin (AeL) nanopores allow direct single nucleotide discrimination of very short oligonucleotides (≤10 nt) without labelling, which shows great potential for DNA sensing. To achieve real applications, one major obstacle of AeL is its poor capture ability of long single-stranded DNA (ssDNA, >10 nt). Here, we have proposed a novel and robust strategy for the electrostatic focusing of long ssDNA into a lithium-chloride (LiCl)-active AeL. By using this method, for the first time we have demonstrated AeL detection of ssDNA longer than 100 nt. Due to screening more negative charges, LiCl improves AeL capture ability of long ssDNA (i.e. 60 nt) by 2.63- to 10.23-fold compared to KCl. Further calculations and molecular dynamics simulations revealed that strong binding between Li+ and the negatively charged residue neutralized the AeL, leading to a reduction in the energy barrier for ssDNA capture. These findings facilitate the future high-throughput applications of AeL in genetic and epigenetic diagnostics.

Based on a plasmon resonance energy transfer (PRET) method, the electrochromic process was imaged in real-time under potential scanning, which achieved the detection of hundreds of molecules on the surface of a single nanoparticle with high time-spatial resolution., Electrochromic materials have attracted increasing attention in the field of smart devices and energy economy due to their excellent reversible chromic properties. Investigating an electrochromic process at the nano-scale is beneficial to the development of functional nano-devices exploiting chromophores. In this study, a new method for real-time imaging of an electrochromic process at the single nanoparticle level is developed based on an ultra-sensitive plasmon resonance energy transfer (PRET) technique. The scattering light intensity of nanoparticles is applied to reveal energy transfer from nanoparticles to chromophores modulated by an electrochromic reaction. This PRET-based technique achieves the detection of hundreds of molecules on the surface of a single nanoparticle. Furthermore, a color-coded amplifying method has been introduced for high-throughput, converting light intensity into easily recognized colors via the Matlab program. Compared with traditional electrochemical imaging techniques, this facile and rapid approach using optical techniques to characterize a real-time electrochemical process significantly enhances detection sensitivity, time and spatial resolution. Notably, the obtained electrochromic behavior of chromophores on a single nanoparticle is in good agreement with the simulated cyclic voltammetry (CV) curves on a nano-electrode. Therefore, this study provides a promising way to simultaneously monitor electrochromic reactions on single nano-electrodes with high-throughput.