Spectroelectrochemical studies of bilayers of phospholipids in gel and liquid state on Au(111) electrode surface.

Differential capacity, chronocoulometry and Polarization Modulation Fourier Transform Infrared Reflection Absorption Spectroscopy (PM FTIRRAS) were employed to investigate spreading of small unilamellar vesicles (SUVs) of DOPC and DMPC onto a Au(111) electrode surface. The electrochemical experiments demonstrated that vesicles fuse onto the electrode surface and at E>-0.5V (SSCE) or at charge densities -10-0.5 V (SSCE), the tilt angle increases to approximately 42 degrees. The increase of the tilt angle is discussed in terms of a change in the packing of the polar head of the phospholipids molecules in the bilayer adsorbed at the electrode surface.

Polarization modulation infrared reflection absorption spectroscopy investigations of thin silica films deposited on gold. 2. Structural analysis of a 1,2-dimyristoyl-sn-glycero-3-phosphocholine bilayer.

In this paper we report on the structural analysis of bilayers of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) using polarization modulation infrared reflection absorption spectroscopy (PM IRRAS). The lipid bilayers were formed on SiO2|Au and Au surfaces using the Langmuir-Blodgett and Langmuir-Schaeffer techniques. As we showed in part 1 (Zawisza, I.; Wittstock, G.; Boukherroub, R.; Szunertis, S. Langmuir 2007, 23, 9303-9309), SiO2 layers of 7 nm thickness, synthesized by plasma-enhanced chemical vapor deposition on 200 nm thick gold covered glass slides, allow PM IRRAS investigations. Only minor changes in the order and structure of the lipid bilayer are observed when deposited on SiO2|Au and Au surfaces. The choline moiety in the leaflet directed toward the SiO2 surface exists in trans conformation and shows a tilt of 28 degrees with the surface normal of the CN bond. On the silica surface in the second leaflet directed toward air and in two layers deposited on the Au surface, trans and gauche isomers of the choline moiety are present and the tilt of the CN bond increases to 55 degrees with respect to the surface normal. The order and molecular orientation in the DMPC bilayers on SiO2 and Au surfaces are not affected by time. The analysis of the phosphate stretching mode on the Au surface shows slight dehydration of this group and reorientation of the phosphate moiety.

Inkjet-printed thiol self-assembled monolayer structures on gold: quality control and microarray electrode fabrication.

Laterally structured, self-assembled monolayers (SAMs) of different thiols (HS-R-X, R = (CH 2) 3-16, X = -CH 3, -COOH, -NH 2) on gold have been prepared by inkjet printing. The printer is a modified, low-cost desktop printer (Epson Stylus Photo R200), the ink is a 1 mM solution of the thiol in ethanol/glycerol (6:1). The quality of inkjet-printed large area SAMs obtained in this study is between that of a layer self-assembled from a thiol solution and that obtained by soft lithography, according to cyclic voltammetry, electrochemical impedance spectroscopy, scanning electrochemical microscopy (SECM), and polarization-modulated Fourier transform infrared reflection-absorption spectroscopy (PM IRRAS). For the first time, simultaneous printing of two different thiols in a single print job as an alternative to sequential printing and backfilling is demonstrated. The smallest structures consisting of conductive disks of 40 microm diameter were analyzed as single spots by SECM and as random array electrodes with different average disk-disk distance. Conductive band electrodes with variable bandwidth (300 microm to 1 cm) are presented, as well as a pH switchable band structure. As compared to stamping, inkjet printing allows for simultaneous multiple thiol printing in a single print job with the resolution limited only by the droplet size and the precision of the translation stage.

Microelectrochemical modulation of micropatterned cellular environments.

Patterned cell cultures obtained by microcontact printing have been modified in situ by a microelectrochemical technique. It relies on lifting cell-repellent properties of oligo(ethylene glycol)-terminated self-assembled monolayers (SAMs) by Br2, which is produced locally by an ultramicroelectrode of a scanning electrochemical microscope (SECM). After Br2 treatment the SAM shows increased permeability and terminal hydrophobicity as characterized by SECM approach curves and contact angle measurements, respectively. Polarization-modulation Fourier transform infrared reflection-absorption spectroscopic (PM FTIRRAS) studies on macroscopic samples show that the Br2 treatment removes the oligo(ethelyene glycol) part of the monolayer within a second time scale while the alkyl part of the SAM degrades with a much slower rate. The lateral extension of the modification can be limited because heterogeneous electron transfer from the gold support destroys part of the electrogenerated Br2 once the monolayer is locally damaged in a SECM feedback configuration. This effect has been reproduced and analyzed by exposing SAM-modified samples to Br2 in the galvanic cell Au|SAM|5 microM Br2 + 0.1 M Na2SO4||10 microM KBr + 0.1 M Na2SO4|Au followed by an PM FTIRRAS characterization of the changes in the monolayer system.

Application of thin titanium/titanium oxide layers deposited on gold for infrared reflection absorption spectroscopy: structural studies of lipid bilayers.

Ultrathin titanium layers when deposited on the surface of gold can be successfully applied for infrared reflection absorption spectroscopy (IRRAS) investigations. It was shown that the reflectivity, the phase shift, and the mean square electric field of the p- and s-polarized IR radiation in up to 20 nm thick titanium layers covered with a 3-4 nm thick layer of native oxide are comparable to those of the air/gold interface. The surface selection rule is fulfilled. Thus, qualitative and quantitative analysis of 1,2-dimyristoyl- sn-glycero-3-phosphocholine (DMPC) bilayers transferred in liquid expanded (LE) and liquid condensed (LC) states can be performed. Differences are found in the hydration state and molecular arrangement of the two investigated bilayers. In the DMPC bilayer in the LE state, the C-N bond in the positively charged choline moiety is inclined by approximately 70 degrees toward the surface of the negatively charged titanium substrate. In the phosphate moiety, the in-plane vector of the O-P-O group makes a small angle of approximately 15 degrees to the surface normal. This open structure of the lipid molecule corresponds to the B crystal structure of the DMPC molecule and provides space for strong hydration of the polar headgroup. In the DMPC bilayer in the LC state, the intermolecular distances are reduced; the C-N bond of the choline group makes a smaller angle to the surface normal, and the in-plane vector of the O-P-O group in the phosphate moiety displays a larger tilt. The degree of hydration is reduced. The arrangement of the polar headgroup region corresponds to the A crystal structure of the DMPC molecule.

Potential-driven structural changes in Langmuir-Blodgett DMPC bilayers determined by in situ spectroelectrochemical PM IRRAS.

Combined Langmuir-Blodgett vertical withdrawing and Langmuir-Schaefer horizontal touch (LB-LS) methods were employed to transfer DMPC bilayers onto a Au(111) electrode surface. Charge density measurements and photon polarization modulation infrared reflection absorption spectroscopy were employed to investigate electric field induced changes in the structure of the bilayer. The results show that the physical state and the molecular arrangement found in the monolayer at the air-water interface is to a large extent preserved in the bilayer formed by the LB-LS method. This approach provides an opportunity to produce supported bilayers with a well-designed architecture. The properties of the bilayer formed by the LB-LS method were compared to the properties of the bilayer produced by spontaneous fusion of unilamellar vesicles investigated in an earlier study (Bin, X.; Zawisza, I.; Lipkowski, J. Langmuir 2005, 21, 330-347). The tilt angles of the acyl chains are much smaller in the bilayer formed by the LB-LS method and are closer to the angles observed for vesicles and stacked hydrated bilayers. The tilt angles of the phosphate and choline groups are also smaller and are characteristic of an orientation in which the area per DMPC molecule is small. The electric field induced changes of these angles are also less pronounced in the bilayer formed by the LB-LS method. We have shown that these differences are a result of the higher packing density of the phospholipid molecules in the bilayer formed by the LB-LS method.

PM IRRAS investigation of thin silica films deposited on gold. Part 1. Theory and proof of concept.

Polarization modulation infrared reflection absorption spectroscopy (PM IRRAS) was successfully used for the first time to characterize an optically transparent thin oxide film. SiO2 layers of 7 nm thickness were synthesized by plasma enhanced chemical vapor deposition (PECVD) on 200 nm thick gold covered glass slides. Despite the fact that silica is transparent and absorptive to IR radiation when deposited in the form of thin films on a gold surface, it preserves the high metallic reflectivity for the IR light. At grazing angles of incidence of the IR beam, the enhancement of the normal component of the electric field at the interface is comparable to that of Au alone. In addition, the analysis of the structure of a 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid monolayer deposited using the Langmuir-Blodgett technique is demonstrated.

Electrochemical and PM-IRRAS studies of the effect of the static electric field on the structure of the DMPC bilayer supported at a Au(111) electrode surface.

Differential capacity, charge density measurements, and polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) were employed to study the fusion of small unilamellar vesicles of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) on a Au(111) electrode surface. The differential capacity and charge density data showed that the vesicles fuse onto the gold surface at charge densities between -10 microC/cm(2) < sigma(M) < 10 microC/cm(2) to form a bilayer. When sigma(M) < -10 microC/cm(2), the film is detached from the surface but it remains in close proximity to the surface. PM-IRRAS experiments provided IR spectra for the bilayer in the adsorbed and the desorbed state. Ab initio normal coordinate calculations were performed to assist interpretation of the IR spectra. The IR bands were analyzed quantitatively, and this analysis provided information concerning the conformation and orientation of the acyl chains and the polar head region of the DMPC molecule. The orientation of the chains, hydration, and conformation of the headgroup of the DMPC molecule strongly depend on the electrode potential.

Layer by layer characterization of 1-octadecanol films on a Au(111) electrode surface. "In situ" polarization modulation infrared reflection absorption spectroscopy and electrochemical studies.

Bilayers of 1-octadecanol were transferred from the gas-solution interface onto a Au(111) electrode surface using either the horizontal touch or the Langmuir-Blodgett (LB) technique. The horizontal touch method gives an X-type film in which the molecule in the bottom layer is oriented with the tail toward the metal and the head toward the tail of the molecule in the top layer (head to tail orientation). The LB technique allows one to build not only X but also a Y bilayer, in which the molecules in the two leaflets are oriented tail to tail with the polar heads facing the metal and the solution. The properties of these films were characterized by electrochemistry and polarization modulation infrared reflection absorption spectroscopy (PM IRRAS). The freshly prepared X bilayer is defect free and is characterized by a low capacity of 0.84 microF cm(-2). The bilayer may be desorbed from the electrode surface at E < -0.6 V and readsorbed again at more positive potentials. The potential-driven readsorption gives a Y film with a higher capacity of 2.2 microF cm(-2). The IR studies of a mixed bilayer composed of one hydrogenated leaflet and a second deuterated leaflet showed that 1-octadecanol molecules assume a more vertical orientation in the leaflet facing the solution than in the leaflet in contact with the metal. Upon desorption of the X film, the molecules in the bottom layer flip-flop and the readsorbed film assumes the Y structure. Smaller tilt angles are observed in the X film as opposed to the Y film.