Chiral Sum Frequency Generation Spectroscopy Detects Double-Helix DNA at Interfaces.

Many DNA-based technologies involve the immobilization of DNA and therefore require a fundamental understanding of the DNA structure-function relationship at interfaces. We present three immobilization methods compatible with chiral sum frequency generation (SFG) spectroscopy at interfaces. They are the "anchor" method for covalently attaching DNA on a glass surface, the "island" method for dropcasting DNA on solid substrates, and the "buoy" method using a hydrocarbon moiety for localizing DNA at the air-water interface. Although SFG was previously used to probe DNA, the chiral and achiral SFG responses of single-stranded and double-stranded DNA have not been compared systemically. Using the three immobilization methods, we obtain the achiral and chiral C-H stretching spectra. The results introduce four potential applications of chiral SFG. First, chiral SFG gives null response from single-stranded DNA but prominent signals from double-stranded DNA, providing a simple binary readout for label-free detection of DNA hybridization. Second, with heterodyne detection, chiral SFG gives an opposite-signed spectral response useful for distinguishing native (D-) right-handed double helix from non-native (L-) left-handed double helix. Third, chiral SFG captures the aromatic C-H stretching modes of nucleobases that emerge upon hybridization, revealing the power of chiral SFG to probe highly localized molecular structures within DNA. Finally, chiral SFG is sensitive to macroscopic chirality but not local chiral centers and thus can detect not only canonical antiparallel double helix but also other DNA secondary structures, such as a poly-adenine parallel double helix. Our work benchmarks the SFG responses of DNA immobilized by the three distinct methods, building a basis for new chiral SFG applications to solve fundamental and biotechnological problems.

Molecular Transport across Oil-Brine Interfaces Impacts Interfacial Tension: Time-Effects in Buoyant and Pendant Drop Measurements.

The buoyant drop method is a ubiquitous tool for addressing phenomena at the liquid-liquid interface via the determination of the interfacial tension (IFT) between two immiscible phases. Here, the focus is on how electrolytes (in an aqueous phase) and carboxylic acids (in a decane phase) impact the interfacial layer between the two phases. The IFT measurement provides a single number, which is not fulfilling when it comes to deducing information about a complex multiparameter system. Furthermore, the temporal evolution of IFT does not always reach a steady-state value on a time scale, which is realistic to use for comparative studies. We have investigated the temporal evolution of IFT in a series of experiments with varying compositions of the decane-carboxylic acid phase and the brine phase. The results show that there are at least two opposing effects in play. For water-soluble acids, the IFT initially increases with time until a turnover point is reached from where there is a gradual decay. The IFT at the turnover point is close to that of the pure water-decane system. For a poorly water-soluble acid, the IFT shows a much smaller increase and the turnover happens much faster. For a water-soluble acid, there is a high degree of sensitivity toward the electrolyte; it determines the position (in time) of the IFT peak and the steepness of the subsequent decay. Now, if the phases are reversed, that is, by placing a drop of brine in the decane-surfactant phase, the IFT decreases with time regardless of the acid and with little impact of the electrolyte and its concentration in the brine. We propose an explanation for the observed behavior (supported by COSMO-RS calculations), which is based on diffusion in and out of the two phases, solubility, and interfacial reactivity (i.e., aggregation between electrolytes and carboxylic acids).