Sum Frequency Generation Results Research Articles

Interface structure of escin at air–water interface probed by sum frequency generation spectroscopy

AbstractSaponins are natural amphiphilic surfactants with high surface activity and unique surface properties at the air–water interfaces. We have applied sum frequency generation (SFG) vibrational spectroscopy to study escin, a kind of saponins that is extracted from chestnut (Aesculus hippocastanum), which adsorbed at water surfaces as a function of the solution concentrations, and have characterized the net orientation of the hydrophobic aglycone moieties at the water interfaces by varying the solution concentration of the escin. SFG results of escin adsorbed water surfaces demonstrate that the hydrophobic aglycone moieties show nearly standing‐up configuration above the critical micelle concentration (cmc), while its preferred orientation is nearly parallel to the surface below cmc. In addition to the changes in the CH stretching band intensities from the escin adsorbed interfaces, we found that the structural arrangement of the escin molecules at the air–water interface is also a function of solution pH and clarified the correlation between the ionic state of the solution and the structure.

Investigation of the Atmospheric Moisture Effect on the Molecular Behavior of an Isocyanate-Based Primer Surface.

A primer coating is engineered to facilitate compatibility between products like adhesives, sealants, and potting compounds and targeted substrates. Prolonged exposure of isocyanate-based primer surfaces to the environment is known to negatively affect the interfacial adhesion between itself and the products subsequently applied on top of it. However, the molecular behavior behind this observed phenomenon remained to be further investigated. In this study, sum frequency generation (SFG) vibrational spectroscopy, a nonlinear optical spectroscopic technique, was applied to study the surface of an isocyanate-based primer exposed to different environments at the molecular level. Atmospheric moisture was considered to be a potential factor in impairing the adhesion performance of the primer, and thus, time- and humidity-dependent experiments were executed to monitor the molecular behavior at the primer surface using SFG. In addition, 180° peel testing experiments were conducted to measure the adhesion properties of primers after being exposed to the corresponding conditions to correlate to SFG results and establish a chemical structure-macroscopic performance relationship. This study on the changes at the primer surface in different environments with varied humidity levels as a function of time aims to provide an in-depth understanding of the moisture effect on isocyanate-based primers. These learnings may also be helpful toward exploring a broader range of coatings and surface layers and improving customer product use guidelines.

Interfacial Behavior of Flux Residues and Its Impact on Copper/Underfill Adhesion in Microelectronic Packaging

AbstractIn the micro-electronics industry, flux treatment on copper is an important procedure to ensure maximum adhesion to copper. In this research, sum frequency generation (SFG) vibrational spectroscopy has been applied to investigate molecular behavior of flux molecules at buried copper and underfill (UF) interfaces in situ. SFG is a second-order nonlinear optical spectroscopic technique, which can provide molecular structural information of surfaces and interfaces with a submonolayer interface specificity. Two model fluxes, adipic acid and phenylacetic acid, and a commercial no-clean flux were examined. Without washing or heating the surface of flux-treated copper, the buried interfaces between copper (treated with flux) and UF are dominated by ordered epoxy in UF. Washing or heating the surface of flux-treated copper leads to disordered copper/UF interfaces, greatly increasing the adhesion. The buried interfacial structures obtained from SFG results are well correlated to the adhesion strengths measured using the lap shear testing method. This research demonstrates the importance of the washing or heating steps of substrate surfaces to increase the interfacial adhesion.

Effects of ball milling on the structure of cotton cellulose

Cellulose is often described as a mixture of crystalline and amorphous material. A large part of the general understanding of the chemical, biochemical and physical properties of cellulosic materials is thought to depend on the consequences of the ratio of these components. For example, amorphous materials are said to be more reactive and have less tensile strength but comprehensive understanding and definitive analysis remain elusive. Ball milling has been used for decades to increase the ratio of amorphous material. The present work used 13 techniques to follow the changes in cotton fibers (nearly pure cellulose) after ball milling for 15, 45 and 120 min. X-ray diffraction results were analyzed with the Rietveld method; DNP (dynamic nuclear polarization) natural abundance 2D NMR studies in the next paper in this issue assisted with the interpretation of the 1D analyses in the present work. A conventional NMR model’s paracrystalline and inaccessible crystallite surfaces were not needed in the model used for the DNP studies. Sum frequency generation (SFG) spectroscopy also showed profound changes as the cellulose was decrystallized. Optical microscopy and field emission-scanning electron microscopy results showed the changes in particle size; molecular weight and carbonyl group analyses by gel permeation chromatography confirmed chemical changes. Specific surface areas and pore sizes increased. Fourier transform infrared (FTIR) and Raman spectroscopy also indicated progressive changes; some proposed indicators of crystallinity for FTIR were not in good agreement with our results. Thermogravimetric analysis results indicated progressive increase in initial moisture content and some loss in stability. Although understanding of structural changes as cellulose is amorphized by ball milling is increased by this work, continued effort is needed to improve agreement between the synchrotron and laboratory X-ray methods used herein and to provide physical interpretation of the SFG results.

Dopant-Dependent SFG Response of Rhenium CO2 Reduction Catalysts Chemisorbed on SrTiO3 (100) Single Crystals

Monolayers of fac-Re(4,4′-dicarboxyl-2,2′-bipyridine)(CO)3Cl (ReC0A) complexes, covalently bound to TiO2-terminated SrTiO3 (100) (STO) single crystals, were studied by both vibrational Sum Frequency Generation (SFG) spectroscopy and Density Functional Theory (DFT). Homodyne-detected SFG results show that the SFG spectral shape of ReC0A on Nb-doped STO surfaces is almost opposite to that of ReC0A on undoped or Fe-doped STO surfaces, which we assign to a phase change of the SFG response of STO induced by the Nb rather than a change in orientation of the molecule. DFT calculations show that the doping with Nb populates the conduction band of the system, giving a metallic nature to the otherwise insulator STO substrate. This change in electronic structures of the substrate leads to a big nonresonant SFG response and thus interference with the spectra. The orientation of the ReC0A on all three substrates was found to be tilted by ∼45–50°, with a bidentate binding geometry and the Cl atom facing the surface. Th...

PH-Dependent Inversion of Hofmeister Trends in the Water Structure of the Electrical Double Layer.

Specific ion effects (SIEs) are known to influence the acid/base behavior of silica and the interfacial structure of water, yet evidence of the effect of pH on SIEs is lacking. Here broadband vibrational sum frequency generation (SFG) spectroscopy was used to study SIEs on the water structure at the electrical double layer (EDL) of silica as a function of pH and monovalent cation identity from pH 2-12 at 0.5 M salt concentration. SFG results indicate a direct Hofmeister series of cation adsorption at pH 8 (Li+ < Na+ < K+ < Cs+), with an inversion in this series occurring at pH > 10. In addition, an inversion in SFG intensity trends also occurred at pH < 6, which was attributed to contributions from asymmetric cation hydration and EDL overcharging. The highly pH-dependent SIEs for silica/water have implications for EDL models that often assume pH-independent parameters.

Identifying the selectivity of antimicrobial peptides to cell membranes by sum frequency generation spectroscopy.

Cationic amphiphilic peptides have been engineered to target both Gram-positive and Gram-negative bacteria while avoiding damage to other cell types. However, the exact mechanism of how these peptides target, bind, and disrupt bacterial cell membranes is not understood. One specific peptide that has been engineered to selectively capture bacteria is WLBU2 (sequence: RRWVRRVRRWVRRVVRVVRRWVRR). It has been suggested that WLBU2 activity stems from the fact that when interacting with bacterial cell membranes the peptide assumes an α-helical structure and inserts itself into the membrane. Alternatively, in the presence of mammalian cell membranes, the peptide assumes an inert β-sheet structure. To test this hypothesis, the authors applied sum frequency generation (SFG) spectroscopy and surface tensiometry to identify the structure of WLBU2 as it interacts with model lipid monolayers that mimic mammalian and bacterial cell membranes. Model mammalian cell membranes were built upon zwitterionic 1,2-dipalmitoyl-sn-glycero-3-phosphocholine lipids while bacterial cell membranes were constructed with negatively charged 1,2-dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol) lipids. Observed changes in surface pressure at the peptide-lipid-air interface demonstrate that the peptide has a clear binding preference toward negatively charged bacteria-like lipids. The structure of both the lipids and peptides were characterized by SFG spectra collected at the monolayer interface. Changes in monolayer structure as the peptide binds were observed by tracking the intensities of SFG vibrational modes related to the acyl chains within the lipids. Peptide structures when bound to both types of lipids were determined by SFG spectra collected within the amide I vibrational band. The SFG spectra of WLBU2 interacting with the model mammalian lipid monolayer contain two peaks near 1642 and 1678 cm-1 indicative of an inactive β-sheet structure. SFG spectra collected from the peptide bound to a bacteria-like lipid monolayer contains just a single peak near 1651 cm-1 which corresponds to an active α-helix structure. Combined, the tensiometry and SFG results demonstrate that WLBU2 both possesses a higher binding affinity toward and is in an active α-helix structure when bound to bacterial cell membranes.

Vibrational spectra and molecular dynamics of hydrogen peroxide molecules at quartz/water interfaces

The influence of H2O2 on the water vibration at quartz interface was examined using sum-frequency generation (SFG) spectroscopy, and the effect of H2O2 concentration has been systematically studied. Further, the number density and radical distribution of water molecules, H2O2 molecules, and quartz surface silanol groups were calculated using molecular dynamics (MD) simulation to provide molecular level interpretation for the SFG spectra. It is concluded from this study that the hydrogen peroxide molecules prefers to donate H-bonds to the in-plane silanol groups rather than accepting H-bonds from out-of-plane silanol groups, as evidenced by the strengthening of the peak located at 3400 cm−1 assigned to “liquid-like” hydrogen-bonding network. The SFG results have been supported by the MD calculation results, which demonstrate that the relative intensity of the peak located at 3400 cm−1 to that of located at 3200 cm−1 increases monotonously with the increase in the number of hydrogen peroxide in the first hydration shell of silanol.

Interfacial Water Structure at Surfactant Concentrations below and above the Critical Micelle Concentration as Revealed by Sum Frequency Generation Vibrational Spectroscopy

Interfacial water in close proximity to an adsorbed surfactant layer plays an important role in many areas. Here, we systematically investigate the interfacial water structure at the adsorption layer of cetyltrimethylammonium bromide (CTAB) from low concentrations to its critical micelle concentration. Our sum frequency generation (SFG) spectroscopy results show that, with increasing CTAB concentration, the water SFG signals first increase, reaching maximum intensities at 0.1 mM, and then drop, whereas the ppp SFG signals of the terminal-methyl asymmetric stretch reach maximum intensities and saturate at 1 mM, which is the critical micelle concentration of CTAB. Our analysis reveals that the interfacial water layer adopts the most orderly arrangement when the interfacial potential of the adsorption layer reaches saturation and not at the surfactant concentration of adsorption saturation. Most importantly, our SFG data provide direct evidence for the antagonistic effects of the interfacial potential and thickness compression of the electrostatic field of the surfactant adsorption layer, leading to the strongest water SFG signals at 0.1 mM. Below 0.1 mM, the increased interfacial potential can have a pronounced effect on the increase of the overall dipole moment of the interfacial water layers. Above 0.1 mM, the decreased Debye screening length significantly reduces the water dipole moment. Finally, the newly proposed adsorption model cannot explain our SFG results.

Surface plasma treatment effects on the molecular structure at polyimide/air and buried polyimide/epoxy interfaces

Polyimides are widely used as chip passivation layers and organic substrates in microelectronic packaging. Plasma treatment has been used to enhance the interfacial properties of polyimides, but its molecular mechanism is not clear. In this research, the effects of polyimide surface plasma treatment on the molecular structures at corresponding polyimide/air and buried polyimide/epoxy interfaces were investigated in situ using sum frequency generation (SFG) vibrational spectroscopy. SFG results show that the polyimide backbone molecular structure was different at polyimide/air and polyimide/epoxy interfaces before and after plasma treatment. The different molecular structures at each interface indicate that structural reordering of the polyimide backbone occurred as a result of plasma treatment and contact with the epoxy adhesive. Furthermore, quantitative orientation analysis indicated that plasma treatment of polyimide surfaces altered the twist angle of the polyimide backbone at corresponding buried polyimide/epoxy interfaces. These SFG results indicate that plasma treatment of polymer surfaces can alter the molecular structure at corresponding polymer/air and buried polymer interfaces.

Molecular interactions between amantadine and model cell membranes.

Sum frequency generation (SFG) vibrational spectroscopy was applied to study molecular interactions between amantadine and substrate supported lipid bilayers serving as model cell membranes. Both isotopically asymmetric and symmetric lipid bilayers were used in the research. SFG results elucidated how the water-soluble drug, amantadine, influenced the packing state of each leaflet of a lipid bilayer and how the drugs affected the lipid flip-flop process. It is difficult to achieve such detailed molecular-level information using other analytical techniques. Especially, from the flip-flop rate change of isotopically asymmetric lipid bilayer induced by amantadine, important information on the drug-membrane interaction mechanism can be derived. The results show that amantadine can be associated with zwitterionic PC bilayers but has a negligible influence on the flip-flop behavior of PC molecules unless at high concentrations. Different effects of amantadine on the lipid bilayer were observed for the negatively charged DPPG bilayer; low concentration amantadine (e.g., 0.20 mM) in the subphase could immediately disturb the outer lipid leaflet and then the lipid associated amantadine molecules gradually reorganize to cause the outer leaflet to return to the original orderly packed state. Higher concentration amantadine (e.g., 5.0 mM) immediately disordered the packing state of the outer lipid leaflet. For both the high and low concentration cases, amantadine molecules only bind to the outer PG leaflet and cannot translocate to the inner layer. The presence of amantadine within the negatively charged lipid layers has certain implications for using liposomes as drug delivery carriers for amantadine. Besides, by using PC or PG bilayers with both leaflets deuterated, we were able to examine how amantadine is distributed and/or oriented within the lipid bilayer. The present work demonstrates that SFG results can provide an in-depth understanding of the molecular mechanisms of interactions between water-soluble drugs and model cell membranes.

Sum frequency generation spectroscopy study of an ionic liquid at a graphene-BaF2 (111) interface.

Sum frequency generation (SFG) vibrational spectroscopy and contact angle measurements of an ionic liquid, 1-butyl-3-methylimidazolium dicyanamide [BMIM][DCA], at solid-liquid interfaces are reported. Bare solid single crystal BaF2 (111) surface, a single and few layer graphene-coated BaF2 (111) surface are used as the solid substrates. The SFG results indicate that both [BMIM](+) and [DCA](-) can be detected specifically on the graphene-coated BaF2 (111) surface, without coating only [DCA](-) are observed. [DCA](-) anions are attracted to the positively charged BaF2 (111) surface and occupy the first layer at the solid-liquid interface. The graphene coating shields the charged crystal surface and allows both cations and anions to exist at the interface. Furthermore, increase in the contact angle of BaF2 surface after graphene layers deposition suggests that the graphene coating lowers the surface energy.

Hysteresis Effects in the In Situ SFG and Differential Capacitance Measurements on Metal Electrode/Ionic Liquids Interface

Potential dependent electric double layer (EDL) structure of ionic liquids on a polycrystalline Pt electrode has been studied by in-situ IR-visible sum frequency generation (SFG) spectroscopy and differential capacitance measurements. Within the electrochemical window (EW), potential dependent adsorption/desorption processes of the ions have been monitored. Both SFG and capacitance results indicate that significant hysteresis occurs during the adsorption/desorption processes on the Pt surface, while the EDL structure is highly reversible at the potential close to the limit of the EW. The unique hysteresis effect for ionic liquids on electrode surface should be seriously considered in electrochemical applications of ionic liquids.

Optical Sum Frequency Generation Image of Rice Grains

We observed optical sum frequency generation (SFG) images of cross-sections of glutinous rice grains, in order to test a possibility of the SFG microscopy as a tool for monitoring polysaccharide species in rice grains. The SFG response in the CH vibration range was the most intense in the crush cell layer at the edge of the endosperm adjacent to the embryo probably due to optical reflection and scattering effectby the rugged dielectric structure of the crush cell layer. The SFG spectra as a function of the infrared wavelength depended on the measurement position in the endosperm. The SFG results were compared with those by Raman and infrared spectroscopies for the same samples.

Observations of Interfacial Population and Organization of Surfactants with Sum Frequency Generation and Surface Tension

The complementary interface-sensitive methods of sum fre- quency generation (SFG) and surface tension have been used to investigate surfactants at the air/water interface. SFG yields information on the density and orientation of surfactants at the interface, whereas surface tension yields information on the surface excess of surfactants, which, however, is not their interfacial density. The rapid rise in the SFG signal to its maximum value at 1 mM, which remained essentially constant thereafter, was attributed to the formation of large surfactant domains. Above 1 mM, the domains, all of which have the same density, dominated the SFG signal. As a consequence, the SFG signalremainedessentiallyconstant.Thesurfaceexcesshadanequallyrapidrise to its maximum value at 1 mM, which remained constant until reaching a concentration of 3 mM, where it dropped to a zero value, indicating that a full surfactant monolayer had been formed. This coincides with the formation of bulk micelles, which is commonly referred to as the critical micelle concentration. The orientation of the surfactant carbonyl chromophore was obtained from polarization measure- ments of the SFG signal and showed a small change at and above a 1 mM concentration. The SFG results and the surface tension results, though inherently different, were found to be consistent with each other.

Membrane orientation of MSI-78 measured by sum frequency generation vibrational spectroscopy.

Antimicrobial peptides (AMPs) selectively disrupt bacterial cell membranes to kill bacteria whereas they either do not or weakly interact with mammalian cells. The orientations of AMPs in lipid bilayers mimicking bacterial and mammalian cell membranes are related to their antimicrobial activity and selectivity. To understand the role of AMP-lipid interactions in the functional properties of AMPs better, we determined the membrane orientation of an AMP (MSI-78 or pexiganan) in various model membranes using sum frequency generation (SFG) vibrational spectroscopy. A solid-supported single 1,2-dipalmitoyl-an-glycero-3-[phospho-rac-(1-glycerol)] (DPPG) bilayer or 1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (POPG) bilayer was used as a model bacterial cell membrane. A supported 1,2-dipalmitoyl-an-glycero-3-phosphocholine (DPPC) bilayer or a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayer was used as a model mammalian cell membrane. Our SFG results indicate that the helical MSI-78 molecules are associated with the bilayer surface with ∼70° deviation from the bilayer normal in the negatively charged gel-phase DPPG bilayer at 400 nM peptide concentration. However, when the concentration was increased to 600 nM, MSI-78 molecules changed their orientation to make a 25° tilt from the lipid bilayer normal whereas multiple orientations were observed for an even higher peptide concentration in agreement with toroidal-type pore formation as reported in a previous solid-state NMR study. In contrary, no interaction between MSI-78 and a zwitterionic DPPC bilayer was observed even at a much higher peptide concentration (∼12,000 nM). These results demonstrate that SFG can provide insights into the antibacterial activity and selectivity of MSI-78. Interestingly, the peptide exhibits a concentration-dependent membrane orientation in the lamellar-phase POPG bilayer and was also found to induce toroidal-type pore formation. The deduced lipid flip-flop from SFG signals observed from lipids also supports MSI-78-induced toroidal-type pore formation.

Probing the Orientation of Surface-Immobilized Protein G B1 Using ToF-SIMS, Sum Frequency Generation, and NEXAFS Spectroscopy

The ability to orient active proteins on surfaces is a critical aspect of many medical technologies. An important related challenge is characterizing protein orientation in these surface films. This study uses a combination of time-of-flight secondary ion mass spectrometry (ToF-SIMS), sum frequency generation (SFG) vibrational spectroscopy, and near-edge X-ray absorption fine structure (NEXAFS) spectroscopy to characterize the orientation of surface-immobilized Protein G B1, a rigid 6 kDa domain that binds the Fc fragment of IgG. Two Protein G B1 variants with a single cysteine introduced at either end were immobilized via the cysteine thiol onto maleimide-oligo(ethylene glycol)-functionalized gold and bare gold substrates. X-ray photoelectron spectroscopy was used to measure the amount of immobilized protein, and ToF-SIMS was used to measure the amino acid composition of the exposed surface of the protein films and to confirm covalent attachment of protein thiol to the substrate maleimide groups. SFG and NEXAFS were used to characterize the ordering and orientation of peptide or side chain bonds. On both substrates and for both cysteine positions, ToF-SIMS data showed enrichment of mass peaks from amino acids located at the end of the protein opposite to the cysteine surface position as compared with nonspecifically immobilized protein, indicating end-on protein orientations. Orientation on the maleimide substrate was enhanced by increasing pH (7.0-9.5) and salt concentration (0-1.5 M NaCl). SFG spectral peaks characteristic of ordered α-helix and β-sheet elements were observed for both variants but not for cysteine-free wild type protein on the maleimide surface. The phase of the α-helix and β-sheet peaks indicated a predominantly upright orientation for both variants, consistent with an end-on protein binding configuration. Polarization dependence of the NEXAFS signal from the N 1s to π* transition of β-sheet peptide bonds also indicated protein ordering, with an estimated tilt angle of inner β-strands of 40-50° for both variants (one variant more tilted than the other), consistent with SFG results. The combined results demonstrate the power of using complementary techniques to probe protein orientation on surfaces.

Structure of the Cetyltrimethylammonium Surfactant at Fused Silica/Aqueous Interfaces Studied by Vibrational Sum Frequency Generation

Utilizing vibrational sum frequency generation (SFG), we characterized the structure of adsorbed cetyltrimethylammonium chloride (CTAC) at the silica/aqueous interface in the presence of 10 to 500 mM NaCl and as a function of surfactant surface coverage. For low ionic strengths (10 mM NaCl), results indicate that adsorbed aggregates do not change conformation with increasing surface coverage. Instead, the surfactant adsorbs as micelle-like structures at concentrations considerably lower than surface saturation and the CMC. At high ionic strengths (300-500 mM NaCl), the structure of the adlayer is considerably different: The SFG results indicate that for 30 microM bulk CTAC the surfactant packs with fewer gauche defects in the hydrocarbon backbone, which is attributed to reduced Coulomb repulsion between the positively charged surfactant headgroups, and the results also indicate that CTAC adsorbs as monomers at low surface coverage but then rearranges into aggregates at higher surface coverage.

Double layer structure and adsorption/desorption hysteresis of neat ionic liquid on Pt electrode surface — an in-situ IR-visible sum-frequency generation spectroscopic study

The electrochemical interface between a polycrystalline Pt electrode and the ionic liquid 1-butyl-3-methylimidazolium trifluoromethanesulfonate ([bmim][OTf]) has been studied by in-situ IR-visible sum-frequency generation (SFG) spectroscopy. Potential dependent adsorption/desorption processes of OTf anions has been monitored within the electrochemical window. SFG results indicate that the ions form a double layer structure at the interface. Significant adsorption/desorption hysteresis has been observed for the anions on the Pt surface.

Molecular interactions between magainin 2 and model membranes in situ.

In this paper, we investigated the molecular interactions of magainin 2 with model cell membranes using sum frequency generation (SFG) vibrational spectroscopy and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR). Symmetric 1-palmitoyl-2-oleoyl-sn-glycero-3-[Phospho-rac-(1-glycerol)] (POPG) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayers, which model the bacterial and mammalian cell membranes, respectively, were used in the studies. It was observed by SFG that magainin 2 orients relatively parallel to the POPG lipid bilayer surface at low solution concentrations, around 200 nM. When increasing the magainin 2 concentration to 800 nM, both SFG and ATR-FTIR results indicate that magainin 2 molecules insert into the POPG bilayer and adopt a transmembrane orientation with an angle of about 20 degrees from the POPG bilayer normal. For the POPC bilayer, even at a much higher peptide concentration of 2.0 microM, no ATR-FTIR signal was detected. For this concentration on POPC, SFG studies indicated that magainin 2 molecules adopt an orientation nearly parallel to the bilayer surface, with an orientation angle of about 75 degrees from the surface normal. This shows that SFG has a much better detection limit than ATR-FTIR and can therefore be applied to study interfacial molecules with a much lower surface coverage. This magainin 2 orientation study and further investigation of the lipid bilayer SFG signals support the proposed toroidal pore model for the antimicrobial activity of magainin 2.