The interfacial region of aqueous systems also known as the electrical double layer can be characterized on the molecular level with second harmonic and sum-frequency generation (SHG/SFG). SHG and SFG are surface specific methods for isotropic liquids. Here, we model the SHG/SFG intensity in reflection, transmission, and scattering geometry taking into account the spatial variation of all fields. We show that, in the presence of a surface electrostatic field, interference effects, which originate from oriented water molecules on a length scale over which the potential decays, can strongly modify the probing depth as well as the expected intensity at ionic strengths <10 −3 M. For reflection experiments this interference phenomenon leads to a significant reduction of the SHG/SFG intensity. Transmission mode experiments from aqueous interfaces are hardly influenced. For SHG/SFG scattering experiments the same interference leads to an increase in intensity and to modified scattering patterns. The predicted scattering patterns are verified experimentally.

The electrostatic environment of aqueous systems is an essential ingredient for the function of any living system. To understand the electrostatic properties and their molecular foundation in soft, living, and three-dimensional systems, we developed a table-top model-free method to determine the surface potential of nano-and microscopic objects in aqueous solutions. Angle-resolved nonresonant second harmonic (SH) scattering measurements contain enough information to determine the surface potential unambiguously, without making assumptions on the structure of the interfacial region. The scattered SH light that is emitted from both the particle interface and the diffuse double layer can be detected in two different polarization states that have independent scattering patterns. The angular shape and intensity are determined by the surface potential and the second-order surface susceptibility. Calibrating the response with the SH intensity of bulk water, a single, unique surface potential value can be extracted. We demonstrate the method with 80 nm bare oil droplets in water and ∼50 nm dioleoylphosphatidylcholine (DOPC) and dioleoylphosphatidylserine (DOPS) liposomes at various ionic strengths.

The microscopic description of the interface of colloidal particles in solution is essential to understand and predict the stability of these systems, as well as their chemical and electrochemical reactivity. However, this description often relies on the use of simplified electrostatic mean field models for the structure of the interface, which give only theoretical estimates of surface potential values and do not provide properties related to the local microscopic structure, such as the orientation of interfacial water molecules. Here we apply polarimetric angle-resolved second harmonic scattering (AR-SHS) to 300 nm diameter SiO 2 colloidal suspensions to experimentally determine both surface potential and interfacial water orientation as a function of pH and NaCl concentration. The surface potential values and interfacial water orientation change significantly over the studied pH and salt concentration range, whereas zeta-potential (ζ) values remain constant. By comparing the surface and ζ-potentials, we find a layer of hydrated condensed ions present in the high pH case, and for NaCl concentrations ≥1 mM. For milder pH ranges (pH < 11), as well as for salt concentrations <1 mM, no charge condensation layer is observed. These findings are used to compute the surface charge densities using the Gouy–Chapman and Gouy–Chapman–Stern models. Furthermore, by using the AR-SHS data, we are able to determine the preferred water orientation in the layer directly in contact with the silica interface. Molecular dynamics simulations confirm the experimental trends and allow deciphering of the contributions of water layers to the total response.

A three-dimensional-phospholipid monolayer with tunable molecular structure was created on the surface of oil nanodroplets from a mixture of phospholipids, oil, and water. This simple nanoemulsion preparation technique generates an in situ prepared membrane model system with controllable molecular surface properties that resembles a lipid droplet. The molecular interfacial structure of such a nanoscopic system composed of hexadecane, 1,2-dihexadecanoyl-sn-glycero-3-phosphocholine (DPPC), and water was determined using vibrational sum frequency scattering and second harmonic scattering techniques. The droplet surface structure of DPPC can be tuned from a tightly packed liquid condensed phase like monolayer to a more dilute one that resembles the liquid condensed/liquid expanded coexistence phase by varying the DPPC/oil/water ratio. The tunability of the chemical structure, the high surface-to-volume ratio, and the small sample volume make this system an ideal model membrane for biochemical research.

Variations between the inner and outer leaflets of cell membranes are crucial for cell functioning and signaling, drug-membrane interactions, and the formation of lipid domains. Transmembrane asymmetry can in principle be comprised of an asymmetric charge distribution, differences in hydration, specific headgroup/H-bonding interactions, or a difference in the number of lipids per leaflet. Here, we characterize the transmembrane asymmetry of small unilamellar liposomes consisting of zwitterionic and charged lipids in aqueous solution using vibrational sum frequency scattering and second harmonic scattering, label-free methods, specifically sensitive to lipid and water asymmetries. For single component liposomes, transmembrane asymmetry is present for the charge distribution and lipid hydration, but the leaflets are not detectably asymmetric in terms of the number of lipids per leaflet, even though geometrical packing arguments would predict so. Such a lipid transmembrane asymmetry can, however, be induced in binary lipid mixtures under conditions that enable H-bonding interactions between phosphate and amine groups. In this case, the measured asymmetry consists of a different number of lipids in the outer and inner leaflet, a difference in transmembrane headgroup hydration, and a different headgroup orientation for the interacting phosphate groups.

We present a method to perform hyper Rayleigh scattering from aqueous solutions and second harmonic scattering measurements from unlabeled interfaces of liposomes and nanoparticles in dilute solutions. The water and interfacial response can be measured on a millisecond timescale, thus opening up the possibility to measure label-free time dependent transport processes in biological (membrane) systems.

Laboratory for fundamental BioPhotonics (LBP), Institute of Bioengineering (IBI), and Institute of Materials Science (IMX), School of Engineering (STI), and Lausanne Centre for Ultrafast Science (LACUS), Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland(Received 21 October 2016; accepted 27 December 2016; published online 25 January 2017)Solutions of liposomes composed of binary mixtures of anionic dioleoylphosphatidylserine (DOPS) and zwitterionic dioleoylphosphatidylcholine (DOPC) are investigated with label-free angle-resolved (AR) second harmonic scattering (SHS) and electrophoretic mobility measurements. The membrane surface potential is extracted from the AR-SHS response. The surface potential changes from −10 to −145 mV with varying DOPS content (from 0% to 100%) and levels off already at ∼10% DOPS content. The ζ-potential shows the same trend but with a drastically lower saturation value (44 mV). This difference is explained by the formation of a condensed layer of Na + counterions around the outer leaflet of the liposome as predicted by charge condensation theories for polyelectrolyte systems.

The molecular structure of zwitterionic and charged monolayers on small oil droplets in aqueous solutions is determined using a combined second harmonic and sum frequency study. From the interfacial vibrational signature of the acyl chains and phosphate headgroups as well as the response of the hydrating water, we find that zwitterionic and charged lipids with identical acyl chains form remarkably different monolayers. Zwitterionic phospholipids form a closely packed monolayer with highly ordered acyl tails. In contrast, the charged phospholipids form a monolayer with a low number density and disordered acyl tails. The charged headgroups are oriented perpendicular to the monolayer rather than parallel, as is the case for zwitterionic lipids. These significant differences between the two types of phospholipids indicate important roles of phospholipid headgroups in the determination of properties of cellular membranes and lipid droplets. The observed behavior of charged phospholipids is different from expectations based on studies performed on extended planar interfaces, at which condensed monolayers are readily formed. The difference can be explained by nanoscale related changes in charge condensation behavior that has its origin in a different balance of interfacial intermolecular interactions.