Here we report on the use of double-nanohole (DNH) optical tweezers as a label-free and free-solution single-molecule probe for protein–DNA interactions. Using this approach, we demonstrate the unzipping of individual 10 base pair DNA-hairpins, and quantify how tumor suppressor p53 protein delays the unzipping. From the Arrhenius behavior, we find the energy barrier to unzipping introduced by p53 to be 2 × 10⁻²⁰ J, whereas cys135ser mutant p53 does not show suppression of unzipping, which gives clues to its functional inability to suppress tumor growth. This transformative approach to single molecule analysis allows for ultra-sensitive detection and quantification of protein–DNA interactions to revolutionize the fight against genetic diseases.

Recent advances in nanoaperture optical tweezers have enabled studies of single nanoparticles like proteins in label-free, free-solution environments.

We use a double nanohole (DNH) optical trap to quantify the size and concentration of nanoparticles in solution. The time to trap shows a linear dependence with nanosphere size and a −2/3 power dependence with nanosphere concentration, which is in agreement with simple microfluidic considerations. The DNH approach has size-specificity on the order of a few nanometers, which was used to selectively quantify particles of a single size within a heterogeneous solution. By looking at individual trapping events, it is in principle possible to extend this approach to the ultimate limit of a single particle concentration, while also being able to operate at high concentrations in the same configuration. In addition, the DNH trap allows us to hold onto individual particles and thereby study constituents of a heterogeneous mixture. By repeating the trapping measurements on spherical particles of different refractive index, we found that the transmission step that indicates trapping scales empirically with the Clausius–Mossotti factor. This approach may be applied to several sensing applications, such as in the study of virus populations, where concentrations vary over many orders of magnitude.

Optomechanics arises from the photon momentum and its exchange with low-dimensional objects. It is well known that optical radiation exerts pressure on objects, pushing them along the light path. However, optical pulling of an object against the light path is still a counter-intuitive phenomenon. Herein, we present a general concept of optical pulling—opto-thermoelectric pulling (OTEP)—where the optical heating of a light-absorbing particle using a simple plane wave can pull the particle itself against the light path. This irradiation orientation-directed pulling force imparts self-restoring behaviour to the particles, and three-dimensional (3D) trapping of single particles is achieved at an extremely low optical intensity of 10⁻² mW μm⁻². Moreover, the OTEP force can overcome the short trapping range of conventional optical tweezers and optically drive the particle flow up to a macroscopic distance. The concept of self-induced opto-thermomechanical coupling is paving the way towards freeform optofluidic technology and lab-on-a-chip devices.