Abhay Kotnala

Quantification of High-Efficiency Trapping of Nanoparticles in a Double Nanohole Optical Tweezer

We measure the dynamics of 20 nm polystyrene particles in a double nanohole trap to determine the trap stiffness for various laser powers. Both the autocorrelation analysis of Brownian fluctuations and the trapping transient analysis provide a consistent value of ∼ 0.2 fN/nm stiffness for 2 mW of laser power, which is similar to theoretical calculations for aperture trapping. As expected, the stiffness increases linearly with laser power. This is comparable to the stiffness obtained for conventional optical traps for trapping, but for ten times smaller dielectric particles and less power. This approach will allow us to quantitatively evaluate future aperture-based optical traps, with the goal of studying the folding dynamics of smaller proteins (∼ 10 kDa) and small-molecule interactions.

Double nanohole optical tweezers visualize protein p53 suppressing unzipping of single DNA-hairpins.

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.

Wedge and gap plasmonic resonances in double nanoholes

We study the plasmonic resonances of double nanoholes (DNHs) in metal films. These apertures exhibit the usual gap-mode Fabry-Pérot resonances, where the zeroth order resonance is determined by the waveguide cut-off and the first order resonance shows sensitivity to the film thickness. An additional wedge resonance is observed, which is sensitive to the curvature of the cusps in the DNHs, analogous to the wedge modes of single wedges. While the gap mode intensity increases dramatically with decreasing gap-width, the wedge mode intensity saturates since its field enhancement arises from the curvature of the metal film, like cylindrical Sommerfeld waves. Experimental transmission spectra agree well with finite-difference time-domain simulations showing these separate resonances. The controlled design of these resonances is critical for applications including optical tweezers, nonlinear conversion, sensing and spectroscopy.

Optical tweezers for free-solution label-free single bio-molecule studies

Nanoaperture based trapping has developed as a significant tool among the various optical tweezer systems for trapping of very small particles down to the single nanometer range. The double nanohole aperture based trap provides a method for efficient, highly-sensitive, label-free, low-cost, free-solution single molecule trapping and detection. We use the double nanohole tweezer to understand biomolecular phenomena like protein unfolding, binding, structural conformation of DNA, protein-DNA interactions, and protein small molecule interactions.

New physics and applications of apertures in thin metal films

The nanoplasmonic properties of apertures in metal films have been studied extensively; however, we have recently discovered surprising new features of this simple system with applications to super-focusing and super-scattering. Furthermore, apertures allow for optical tweezers that can hold onto particles of the order of 1 nm; I will briefly highlight our work using these apertures to study protein - small molecule interactions and protein - DNA binding.

Trapping, unfolding, identifying, and binding single proteins using the double-nanohole optical trap

In this paper we describe the double nanohole laser tweezer system used to trap single nanoparticles. We cover the basic theory behind the DNH and what makes it more powerful than traditional laser tweezers commonly used for larger particles. We outline the basic setup used to reliably trap several different types of particles ranging in size from 1 nm to 40 nm. Data from several experiments is shown which displays exactly how a particle is confirmed to be trapped. We will discuss the use of autocorrelation as well as other information that can be extracted from the optical transmission in our setup and how it has been applied to the identification of protein small molecule interactions and protein binding. Other uses of the data collected from our setup will be discussed including the observation of protein folding. Finally we discuss the current developments of the process and its possible uses as a drug discovery tool, a new type of single particle nanopipette and new bio-sensors.