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Lateral flow assays (LFAs) widely deployed for on-site diagnosis have predominantly utilized antibodies as recognition molecules. Antibodies with limited thermal stability deteriorate the performance of the LFA over time. Herein, we demonstrate a stable and robust LFA by utilizing thermally stable peptide-based 12-14 kDa affimers as recognition molecules, in lieu of conventional protein-based antibodies to analyze complex samples with a significantly improved shelf life at room temperature. The model system studied here is that of interleukin-8 (IL8) biomarker for validating the efficacy of the proposed approach, using a pair of affimer probes that demonstrates dual functionality of capturing and reporting. Affimers immobilized on the test zone of LFA serve as capture probes for IL8-affimer-MB complexes. Whereas affimers conjugated with the MBs that enable extraction of IL8 from the sample matrix serve as reporters for visual detection. The MB complexes captured at the test zone resulted in brownish test bands that enable concentration-dependent detection of IL8. The assay yielded sensitive visual detection of IL8 at ng/mL levels (~ 0.1 ng/mL and 1 ng/mL in buffer and human plasma, respectively), within 20 min, using sample volumes of ~ 100 µL. Importantly, the stability of affimer-incorporated LFA improved significantly in contrast to antibody-incorporated LFA over time, even when stored at 4 °C. Therefore, the proposed affimer-based LFA in conjunction with MBs offer stable and reliable detection of biomarkers at clinically relevant concentration ranges in complicated matrices, even without requiring cold storage, hence, offering a promising avenue for on-site diagnosis in resource-limited settings. Nanyang Technological University The Institute for Nanomedicine jointly established between Northwestern University and Nanyang Technological University provided fnancial support

Urinalysis enables non-invasive point-of-care (POC) testing of numerous biomarkers at their physiological and elevated levels, obviating the need for sophisticated equipment or trained personnel. POC urinalysis is used to identify biomarkers that are rich in urine (greater than 1 μM), such as lactate, uric acid, glucose, ions, and adenosine. Urine also contains biomarkers such as small molecules, nucleic acids, neurotransmitters, and drugs in trace amounts (less than 1 μM). These biomarkers are of significant importance for health care monitoring, diagnosis of various disorders (cancer, metabolic diseases, etc.) and illicit drug control (cocaine, steroids, etc.). While POC detection of urinary biomarkers at higher concentration (μM to mM) levels is feasible, direct assaying of biomarkers in nM to fM levels is challenging, as assay responses are typically masked by interferences from the urine sample matrix. This report is a consolidated review of emerging trends and challenges in the POC urinalysis for detecting biomarkers that are less abundant in urine. The sensing mechanisms, analytical device fabrication, discrete and integrated sample pre-treatment procedures for POC assaying of urinary markers in trace amounts are elaborated. Subsequently, the utilization of smart data analytics for facilitating personalized urinalysis is presented. A comprehensive outlook on associated challenges in POC urinalysis of biomarkers in trace amounts is further provided, which would facilitate the advancement of POC urinalysis for a wide range of healthcare applications.

A flow-through colorimetric assay for detection of nucleic acids in plasma is reported. The proposed assay features an array of four polyvinylidene fluoride (PVDF) membranes impregnated with cationic poly (3-alkoxy-4-methylthiophene) (PT) as an optical reporter. The sensing strategy is based on monitoring the changes in optical properties of PT, upon complexation with target nucleic acids in the presence and in the absence of their corresponding complementary peptide nucleic acids (PNAs). As a proof of concept, the proposed methodology is validated using two biomarkers; lung cancer associated microRNA (mir21) and hepatitis B virus DNA (HBV-DNA). The flow-through colorimetric assay enabled detection of mir21 and HBV-DNA in plasma without requiring tedious sample pre-treatment and clean up protocols. Colorimetric responses for mir21 and HBV-DNA were obtained at nanomolar concentrations over five orders of magnitudes (from 1 nM to 10 μM), with a limit of detection of ∼0.6 nM and ∼2 nM in DI water and plasma, respectively. A logic gate system was developed to utilize the colorimetric assay responses as inputs for discrimination of mir21 and HBV-DNA and subsequently to obtain a profile of nucleic acids in samples that exceed respective clinical threshold limits, thereby enabling rapid and point of care (POC) disease diagnosis. Furthermore, the proposed methodology can be utilized for detection of a large number of nucleic acids in plasma by extending the array of PT impregnated membranes incorporated with their corresponding complementary PNAs. Accepted version

Point of care testing (POCT) platforms are analytical devices used for rapid on-site sensing applications. POCT in sports science has attracted significant attention as the markers related to physiological changes and metabolism can be analysed on-site for monitoring an athlete's health, performance, recovery and even for doping control. POCT platforms also possess the potential to facilitate trainers in optimizing the training regimes for preventing injuries, track their nutrition, and could potentially assist sports committees to enforce effective doping control measures during and outside competitions. This review is a consolidated report on emerging trends in POCT platforms in sports science. The methodologies reported thus far that have already been applied or possess a potential to be applied for detection of various sports biomarkers are elaborated. A comprehensive outlook is further provided, which would aid efforts on development of POCT platforms for a wide range of applications. Ministry of Education (MOE) Nanyang Technological University We would like to thank to MOE Tier 1 Grant, Singapore (Grant number 2018-T1-001-079) and Provost Office NTU, Singapore for supporting this work.

Efficient removal of interferents from complex matrices would significantly improve the performance of state of the art dipstick assays. Herein, we evaluate a graphitized carbon black (GCB)-incorporated dipstick, a configuration that has not been explored before, for reliable and facile on-site analysis of complex matrices. Carrot juice, a highly pigmented sample matrix, is chosen for evaluating the retention of interferents within the sorbent-incorporated cleanup pad on the dipstick. A peptide with a specific cleavage site for botulinum neurotoxin A light chain (BoNT/A LC), a model protease for validation of the proposed dipstick assay, is incubated with the test samples containing BoNT/A LC. Subsequently, the BoNT/A LC digested substrate and sample matrix flow vertically through the GCB-deposited cleanup pad within which the matrix interferents are captured, while the substrate, with a minimum of interferents, continues to flow toward a conjugation pad for labelling with Europium particles. Finally, the cleaved and uncleaved substrates flow toward a detection zone, where they bind to the test line producing a pinkish band which is not visible in the absence of GCB incorporation. The dipstick assay yields a LOD of 0.1 nM (5 ng/mL) of BoNT/A LC in carrot juice, within 20 min. The reported approach enables detection of proteases in a wide range of matrices upon incorporation of appropriate sorbents, ultimately aiming to exclude tedious laboratory-based sample pre-treatment protocols. Thus, merging extraction, cleanup, and pre-concentration steps with a sensitive optical detection approach is an attractive strategy for on-site assaying in complex matrices. Graphical abstract.

Development of new detection methodologies and amplification schemes is indispensable for plasmonic biosensors to improve the sensitivity for the detection of trace amounts of analytes. Herein, an ultrasensitive scheme for signal enhancement based on the concept of surface-plasmon-resonance-enhanced light scattering (SP-LS) was validated experimentally and theoretically. The SP-LS of gold nanoparticles' (AuNPs) tags was employed in a sandwich assay for the detection of cardiac troponin I and provided up to 2 orders of magnitude improved sensitivity over conventional AuNPs-enhanced refractometric measurements and 3 orders of magnitude improvement over label-free SPR. Simulations were also performed to provide insights into the physical mechanisms. Refereed/Peer-reviewed

Using single-particle tracking, we investigate the interaction of small unilamellar vesicles (SUVs) that are electrostatically tethered to the freestanding membrane of a giant unilamellar vesicle (GUV). We find that the surface mobility of the GUV-riding SUVs is Brownian, insensitive to the bulk viscosity, vesicle size, and vesicle fluidity but strongly altered by the viscosity of the underlying membrane. Analyzing the diffusional behavior of SUVs within the Saffman-Delbrück model for the dynamics of membrane inclusions supports the notion that the mobility of the small vesicles is coupled to that of dynamically induced lipid clusters within the target GUV membrane. The reversible binding also offers a nonperturbative means for measuring the viscosity of biomembranes, which is an important parameter in cell physiology and function.

A colorimetric Al3+ sensor based on fluorescence recovery of a conjugated copolymer-ATP complex is proposed. An optimized ratio of two polythiophene (PT) monomers is utilized to synthesize copolymer (CP) that yielded maximized colorimetric response for Al3+ in deionized (DI) and tap water. The electrostatic disassembly of CP-ATP upon addition of Al3+ led to an evident visual color change. The lowest concentration of Al3+ for naked eye observation is around 4 μM, which is below the threshold levels in drinking water according to European Economic Community (EEC) standard. Besides, the proposed assay showed a similar response to Al3+ in tap water. The proposed methodology showed selective and sensitive detection for Al3+ in analytically relevant concentration ranges without involving sophisticated instrumentation, illustrating the applicability for on-site drinking water monitoring., MOE (RG 82/12); Provost Office Nanyang Technological University, Singapore

Early detection of toxic proteases in food matrices plays a major role in preventing the occurrence of diseases as well as outbreaks. However, on-site detection of proteases, for instance, botulinum, anthrax and cholera in food matrices remains challenging due to their extremely low lethal dose levels. Here, we report a lateral flow assay (LFA) in a dipstick format for on-site visual detection of proteases in food matrices. The light chain of BoNT serotype A (BoNT/A LC) is used as a model system for validation of the proposed assay using magnetic beads conjugated to a synthetic peptide that provide a specific cleavage site for BoNT/A LC. Magnetic beads serve as both reporters for visual detection and as facilitators for sample clean-up, owing to the efficient magnetic separation protocol adopted. Digestion of the peptide substrate by BoNT/A LC for 5 h followed by the dipstick assay yields a reduction in color intensity of the test line on the dipstick compared to the control line obtained using an un-cleaved peptide substrate. Concentration dependent responses for the assay in carrot juice were obtained with a limit of detection (LOD) of 1 nM/2.5 nM (with/without amplification), also supported by RGB (ΔE) analysis, indicating the potential of the proposed methodology for on-site assaying of proteases in food matrices. Unlike typical affinity-based assays that yield a collective response for the active and inactive forms of the proteases, the proposed functional LFA targets only the active form, thereby enabling a more precise analysis for preventing potential false-positives. The proposed approach could be extended for detection of BoNT serotypes and other proteases in food matrices, upon utilizing appropriate substrates with specific cleavage sites.

International audience; It is of paramount importance to be able to accurately quantify surface coverage of antibodies on gold nanoparticles (AuNP) so as to optimise the sensitivity of AuNP-based immunosensors. Herein, we developed a fluorescence-based method to directly quantify rabbit immunoglobulin G (IgG) used as antibody model bound to AuNP. Rabbit IgG was first labelled with fluorescein-5-isothiocyanate (FITC) prior to conjugation to AuNP via either physisorption or chemisorption. IgG-conjugated AuNP were treated with NaCN to dissolve the AuNP and restore the fluorescence emission that was quenched in the presence of the metallic colloids, followed by quantification of fluorescein by spectrofluorimetry. This direct assay gave about 4 IgG bound to each 15-nm diameter AuNP for both immobilization strategies. This surface coverage value was in good agreement with that determined from the theoretical value calculated from the Localized Surface Plasmon Resonance (LSPR) band shift. For comparison, we also applied two indirect methods based on the quantitation of excess IgG remaining in the supernatant using fluorescence assay or enzyme-linked immunosorbent assay (ELISA). The indirect assays, either fluorescence or ELISA, commonly used to assess the antibody coverage on AuNP, overestimated the IgG surface coverage to a large extent, since up to 3 to 4 times higher coverages were measured. Therefore, the direct fluorescence method reported in this paper appears as a valuable method for quantification of surface coverage of antibody on AuNP.

Conjugated polyelectrolytes (CPEs) have been widely used as reporters in colorimetric assays targeting nucleic acids. CPEs provide naked eye detection possibility by their superior optical properties however, as concentration of target analytes decrease, trace amounts of nucleic acid typically yield colorimetric responses that are not readily perceivable by naked eye. Herein, we report a pixelated analysis approach for correlating colorimetric responses of CPE with nucleic acid concentrations down to 1 nM, in plasma samples, utilizing a smart phone with an algorithm that can perform analytical testing and data processing. The detection strategy employed relies on conformational transitions between single stranded nucleic acid-cationic CPE duplexes and double stranded nucleic acid-CPE triplexes that yield distinct colorimetric responses for enabling naked eye detection of nucleic acids. Cationic poly[N,N,N-triethyl-3-((4-methylthiophen-3-yl)oxy)propan-1-aminium bromide] is utilized as the CPE reporter deposited on a polyvinylidene fluoride (PVDF) membrane for nucleic acid assay. A smart phone application is developed to capture and digitize the colorimetric response of the individual pixels of the digital images of CPE on the PVDF membrane, followed by an analysis using the algorithm. The proposed pixelated approach enables precise quantification of nucleic acid assay concentrations, thereby eliminating the margin of error involved in conventional methodologies adopted for interpretation of colorimetric responses, for instance, RGB analysis. The obtained results illustrate that a ubiquitous smart phone could be utilized for point of care colorimetric nucleic acids assays in complex matrices without requiring sophisticated software or instrumentation., Graphical abstract Image 1, Highlights • A cost effective sensory cartridge and an algorithm compatible with smart phone are developed. • Conjugated polyelectrolyte impregnated paper for detection of nucleic acid. • A pixelated assay to visualize nucleic acid down to concentration of 1 nM.

A volatile organic compounds (VOC) sensor built on SOI-based photonic platform is presented in this paper. The sensor consists of a high-Q factor ring-resonator. It is tested with acetone vapor, achieving a detection limit of 200 ppm. The sensor obtained relative high sensitivity (1.7 pm/1000 ppm), fast response time (∼20 s) and the stability of

In this review, recent advances in the development of electronic detection methodologies based on non-antibody recognition elements such as functional liposomes, aptamers and synthetic peptides are discussed. Particularly, we highlight the progress of field effect transistor (FET) sensing platforms where possible as the number of publications on FET-based platforms has increased rapidly. Biosensors involving antibody-antigen interactions have been widely applied in diagnostics and healthcare in virtue of their superior selectivity and sensitivity, which can be attributed to their high binding affinity and extraordinary specificity, respectively. However, antibodies typically suffer from fragile and complicated functional structures, large molecular size and sophisticated preparation approaches (resource-intensive and time-consuming), resulting in limitations such as short shelf-life, insufficient stability and poor reproducibility. Recently, bio-sensing approaches based on synthetic elements have been intensively explored. In contrast to existing reports, this review provides a comprehensive overview of recent advances in the development of biosensors utilizing synthetic recognition elements and a detailed comparison of their assay performances. Therefore, this review would serve as a good summary of the efforts for the development of electronic bio-sensing approaches involving synthetic recognition elements.

Lipopolysaccharide (LPS) is a toxic inflammatory stimulator released from the outer cell membrane of Gram-negative bacteria, known to be directly related to, for example, septic shock, that causes millions of casualties annually. This number could potentially be lowered significantly if specific, sensitive, and more simply applicable LPS biosensors existed. In this work, we present a facile, sensitive and selective LPS sensor, developed by assembling tetramethylrhodamine-labeled LPS-binding peptides on graphene oxide (GO). The fluorescence of the dye-labeled peptide is quenched upon interaction with GO. Specific binding to LPS triggers the release of the peptide-LPS complex from GO, resulting in fluorescence recovery. This fluorescent turn-on sensor offers an estimated limit of detection of 130 pM, which is the lowest ever reported among all synthetic LPS sensors to date. Importantly, this sensor is applicable for detection of LPS in commonly used clinical injectable fluids, and it enables selective detection of LPS from different bacterial strains as well as LPS on the membrane of living E. coli.

Hizal, Gurkan/0000-0003-1238-362X; Liedberg, Bo/0000-0003-2883-6953, WOS: 000430512200028, PubMed: 29557164, This study reports on a hand-held volatilome analyzer for selective determination of clinically relevant bio-markers in exhaled breath. The sensing platform is based on electrospun polymer nanofiber-multiwalled carbon nanotube (MWCNT) sensing microchannels. Polymer nanofibers of poly(vinylidene fluoride) (PVDF), polystyrene (PS), and poly(methyl methacrylate) (PMMA) incorporated with MWCNT exhibits a stable response to interferences of humidity and CO2 and provides selective deformations upon exposure of exhaled breath target volatilomes acetone and toluene, exhibiting correlation to diabetes and lung cancer, respectively. The sensing microchannels "P1" (PVDF-MWCNT), "P2" (PS-MWCNT), and "P3" (PMMA-MWCNT) are integrated with a microfluidic cartridge (mu-card) that facilitates collection and concentration of exhaled breath. The volatilome analyzer consists of a conductivity monitoring unit, signal conditioning circuitries and a low energy display module. A combinatorial operation algorithm was developed for analyzing normalized resistivity changes of the sensing microchannels upon exposure to breath in the concentration ranges between 35 ppb and 3.0 ppm for acetone and 1 ppb and 10 ppm for toluene. Subsequently, responses of volatilomes from individuals in the different risk groups of diabetes were evaluated for validation of the proposed methodology. We foresee that proposed methodology provides an avenue for rapid detection of volatilomes thereby enabling point of care diagnosis in high-risk group individuals.

This article reports on carbon nanotube/manganese dioxide (CNT-MnO2) composites as electrochemical tags for non-enzymatic signal amplification in immunosensing. The synthesized CNT-MnO2 composites showed good electrochemical activity, electrical conductivity and stability. The electrochemical signal of CNT-MnO2 composites coated glassy carbon electrode (GCE) increased by nearly two orders of magnitude compared to bare GCE in hydrogen peroxide (H2O2) environment. CNT-MnO2 composite was subsequently validated as electrochemical tags for sensitive detection of α-fetoprotein (AFP), a tumor marker for diagnosing hepatocellular carcinoma. The electrochemical immunosensor demonstrated a linear response on a log-scale for AFP concentrations ranging from 0.2 to 100 ng mL(-1). The limit of detection (LOD) was estimated to be 40 pg mL(-1) (S/N=3) in PBS buffer. Further measurements using AFP spiked plasma samples revealed the applicability of fabricated CNT-MnO2 composites for clinical and diagnostic applications.

Surface plasmon field-enhanced fluorescence is reported for the readout of a heterogeneous assay that utilizes low affinity split aptamer ligands. Weak affinity ligands that reversibly interact with target analytes hold potential for facile implementation in continuous monitoring biosensor systems. This functionality is not possible without the regeneration of more commonly used assays relying on high affinity ligands and end-point measurement. In fluorescence-based sensors, the use of low affinity ligands allows avoiding this step but it imposes a challenge associated with the weak optical response to the specific capture of the target analyte which is also often masked by a strong background. The coupling of fluorophore labels with a confined field of surface plasmons is reported for strong amplification of the fluorescence signal emitted from the sensor surface and its efficient discrimination from the background. This optical scheme is demonstrated for time-resolved analysis of chosen model analytes – adenoside and adenosine triphosphate – with a split aptamer that exhibits an equilibrium affinity binding constant between 0.73 and 1.35 mM. The developed biosensor enables rapid and specific discrimination of target analyte concentration changes from low μM to mM in buffer as well as in 10% serum.

We describe the engineering of stable gold nanoparticle (AuNP) bioconjugates for the detection of staphylococcal enterotoxin A (SEA) using localized surface plasmon resonance (LSPR). Two types of AuNP bioconjugates were prepared by covalently attaching anti-SEA antibody (Ab) or SEA to AuNPs. This was achieved by reacting Traut's reagent with lysine residues of both proteins to generate thiol groups that bind to gold atoms on the AuNP surface. These bioconjugates were characterized in-depth by absorption spectroscopy, cryo-transmission electron microscopy, dynamic light scattering, and zeta potential measurements. Their stability over time was assessed after 1 year storage in the refrigerator at 4 °C. Two formats of homogeneous binding assays were set up on the basis of monitoring of LSPR peak shifts resulting from the immunological reaction between the (i) immobilized antibody and free SEA, the direct assay, or (ii) immobilized SEA and free antibody, the competitive assay. In both formats, a correlation between the LSPR band shift and SEA concentration could be established. Though the competitive format did not meet the expected analytical performance, the direct format, the implementation of which was very simple, afforded a specific and sensitive response within a broad dynamic range-nanogram per milliliter to microgram per milliliter. The limit of detection (LOD) of SEA was estimated to equal 5 ng/mL, which was substantially lower than the LOD obtained using a quartz crystal microbalance. Moreover, the analytical performance of AuNP-Ab bioconjugate was preserved after 1 year of storage at 4 °C. Finally, the LSPR biosensor was successfully applied to the detection of SEA in milk samples. The homogeneous nanoplasmonic immunosensor described herein provides an attractive alternative for stable and reliable detection of SEA in the nanogram per milliliter range and offers a promising avenue for rapid, easy to implement, and sensitive biotoxin detection. Sensitive LSPR Biosensing of SEA in buffer and milk using stable AuNP-Antibody bioconjugates Graphical abstract.

Advanced chemometric analysis is required for rapid and reliable determination of physical and/or chemical components in complex gas mixtures. Based on infrared (IR) spectroscopic/sensing techniques, we propose an advanced regression model based on the extreme learning machine (ELM) algorithm for quantitative chemometric analysis. The proposed model makes two contributions to the field of advanced chemometrics. First, an ELM-based autoencoder (AE) was developed for reducing the dimensionality of spectral signals and learning important features for regression. Second, the fast regression ability of ELM architecture was directly used for constructing the regression model. In this contribution, nitrogen oxide mixtures (i.e., N2O/NO2/NO) found in vehicle exhaust were selected as a relevant example of a real-world gas mixture. Both simulated data and experimental data acquired using Fourier transform infrared spectroscopy (FTIR) were analyzed by the proposed chemometrics model. By comparing the numerical results with those obtained using conventional principle components regression (PCR) and partial least square regression (PLSR) models, the proposed model was verified to offer superior robustness and performance in quantitative IR spectral analysis., publishedVersion

International audience; We report on a smartphone spectrometer for colorimetric biosensing applications. The spectrometer relies on a sample cell with an integrated grating substrate, and the smartphone's built-in light-emitting diode flash and camera. The feasibility of the smartphone spectrometer is demonstrated for detection of glucose and human cardiac troponin I, the latter in conjunction with peptide-functionalized gold nanoparticles.

A novel approach for rapid and sensitive detection of matrilysin (MMP-7, a biomarker involved in the degradation of various macromolecules) based on a polypeptide (JR2EC) functionalized reduced graphene oxide (rGO) field effect transistor (FET) is reported. MMP-7 specifically digests negatively charged JR2EC immobilized on rGO, thereby modulating the conductance of rGO-FET. The proposed assay enabled detection of MMP-7 at clinically relevant concentrations with a limit of detection (LOD) of 10 ng/mL (400 pM), attributed to the significant reduction of the net charge of JR2EC upon digestion by MMP-7. Quantitative detection of MMP-7 in human plasma was further demonstrated with a LOD of 40 ng/mL, illustrating the potential for the proposed methodology for tumor detection and carcinoma diagnostic (e.g., lung cancer and salivary gland cancer). Additionally, excellent specificity of the proposed assay was demonstrated using matrix metallopeptidase 1 (MMP-1), a protease of the same family. With appropriate selection and modification of polypeptides, the proposed assay could be extended for detection of other enzymes with polypeptide digestion capability.

Resonance Raman spectroscopy (RRS) often suffers from the large fluorescence background which obscures the much weaker Raman scattering. To address this fundamental problem, a novel Raman substrate ...

A series of alkylthiol compounds were synthesized to study the formation and structure of complex self-assembled monolayers (SAMs) consisting of interchanging structural modules stabilized by intermolecular hydrogen bonds. The chemical structure of the synthesized compounds, HS(CH(2))(15)CONH(CH(2)CH(2)O)(6)CH(2)CONH-X, where X refers to the extended chains of either -(CH(2))(n)CH(3) or -(CD(2))(n)CD(3), with n = 0, 1, 7, 8, 15, was confirmed by NMR and elemental analysis. The formation of highly ordered, methyl-terminated SAMs on gold from diluted ethanolic solutions of these compounds was revealed using contact angle goniometry, null ellipsometry, cyclic voltammetry, and infrared reflection absorption spectroscopy. The experimental work was complemented with extensive DFT modeling of infrared spectra and molecular orientation. New assignments were introduced for both nondeuterated and deuterated compounds. The latter set of compounds also served as a convenient tool to resolve the packing, conformation, and orientation of the buried and extended modules within the SAM. Thus, it was shown that the lower alkyl portion together with the hexa(ethylene glycol) portion is stabilized by the two layers of lateral hydrogen bonding networks between the amide groups, and they provide a structurally robust support for the extended alkyls. The presented system can be considered to be an extension of the well-known alkyl SAM platform, enabling precise engineering of nanoscopic architectures on the length scale from a few to approximately 60 A for applications such as cell membrane mimetics, molecular nanolithography, and so forth.

This work describes the fabrication, characterization, and protein adsorption behavior of charged polymer gradients. The thin gradient films were fabricated by a two-step technique using UV-initiated free-radical polymerization in a reactor with a moving shutter. A homogeneous layer of cationic poly(2-aminoethyl methacrylate hydrochloride) was first formed, followed by a layer of oppositely charged poly(2-carboxyethyl acrylate) with a continuously increasing thickness. Adsorption from protein solutions as well as human blood plasma was investigated by imaging surface plasmon resonance and infrared microscopy. The results showed excessive protein adsorption in the areas where one of the polymers dominated the composition, while there was a clear minimum at an intermediate position of the gradient. The charge of the surface was estimated by direct force measurements and found to correlate well with the protein adsorption, showing the lowest net charge in the same area as the protein adsorption minimum. We therefore hypothesize that a combination of the charged polymers, in the right proportions, can result in a protein-resistant surface due to balanced charges.

A novel approach for miRNA assay using a cationic polythiophene derivative, poly[3-(3′-N,N,N-triethylamino-1′-propyloxy)-4-methyl-2,5-thiophene hydrobromide] (PT), immobilized on a quartz resonator is proposed. The cationic PT enables capturing of all RNA sequences in the sample matrix via electrostatic interactions, resulting in the formation of PT-RNA duplex structures on quartz resonators. Biotinylated peptide nucleic acid (b-PNA) sequences are subsequently utilized for the RNA assay, upon monitoring the PT-RNA-b-PNA triplex formation. Signal amplification is achieved by anchoring avidin coated nanoparticles to b-PNA in order to yield responses at clinically relevant concentration regimes. Unlike conventional nucleic acid assay methodologies that usually quantify a specific sequence of RNA, the proposed approach enables the assay of any RNA sequence in the sample matrix upon hybridization with a PNA sequence complementary to the RNA of interest. As an illustration, successful detection of mir21, (a miRNA sequence associated with lung cancer) is demonstrated with a limit of detection of 400 pM. Furthermore, precise quantification of mir21 in plasma samples is demonstrated without requiring PCR and sophisticated instrumentation.

This contribution describes the synthesis, characterisation and evaluation of a novel biochip technology for the detection of the explosive substance 2,4,6-trinitrotoluene (TNT). Two types of thiols are self-assembled to produce the biochip on gold, namely oligo(ethylene glycol) (OEG)-alkyl thiols terminated with a hydroxyl group and a TNT-analogue (2,4-dinitrobenzene), respectively. Three different TNT-analogues are mixed in various proportions with hydroxyl-terminated OEG-thiols to obtain highly selective and sensitive biochips with a low non-specific binding. The produced self-assembled monolayers (SAMs) are thoroughly characterised with null ellipsometry, contact angle goniometry, infrared reflection absorption spectroscopy (IRAS) and X-ray photoelectron spectroscopy (XPS) and they all meet high standards in terms of molecular conformation, packing and orientation. The biochip is designed to function as a platform for a competitive label-free immunoassay and two real-time transducers – surface plasmon resonance (SPR) and quartz crystal microbalance (QCM) – are used to monitor the dissociation of on-line immobilised monoclonal antibodies produced against TNT. The three TNT-analogues are all potential candidates for the development of a functional biochip, though one of them displayed superior properties in terms of shorter recovery/stabilisation time after antibody immobilisation and a better response/loading capacity ratio. This is particularly evident when using low antigen (TNT-analogue) content in the mixed SAM.

A variety of molecular gradients of alkanethiols with the structure HS-(CH(2))(m)-X (m = 15; X = COOH, CH(2)NH(2), or CH(3)) and oligo(ethylene glycol)-terminated alkanethiols with the structures HS-(CH(2))(15)-CO-NH-Eg(n) (n = 2, 4, or 6), HS-(CH(2))(15)-CO-NH-Eg(2)-(CH(2))(2)-NH-CO-(CH(2))(4)-biotin, and HS-(CH(2))(15)-CO-NH-Eg(6)-CH(2)-COOH were prepared on polycrystalline gold films. These gradients were designed to serve as model surfaces for fundamental studies of protein adsorption and immobilization phenomena. Ellipsometry, infrared spectroscopy, and X-ray photoelectron spectroscopy, operating in scanning mode, were used to monitor the layer composition, gradient profiles, tail group conformation, and overall structural quality of the gradient assemblies. The gradient profiles were found to be 4-10 mm wide, and they increased in width with increasing difference in molecular complexity between the thiols used to form the gradient. The oligo(ethylene glycol) thiols are particularly interesting because they can be used to prepare so-called conformational gradients, that is, gradients that display a variation in oligo(ethylene glycol) chain conformation from all trans on the extreme Eg(2,4) sides, via an amorphous-like phase in the mixing regimes, to helical at the extreme Eg(6) sides. We demonstrate herein a series of experiments where the above gradients are used to evaluate nonspecific binding of the plasma protein fibrinogen, and in agreement with previous studies, the highest amounts of nonspecifically bound fibrinogen were observed on all-trans monolayers, that is, on the extreme Eg(2,4) sides. Moreover, gradients between Eg(2) and a biotinylated analogue have been prepared to optimize the conditions for the immobilization of streptavidin. Ellipsometry and infrared spectroscopy reveal high levels of immobilization over a fairly broad range of compositions in the gradient regime, with a maximum between 50 and 60% of the biotinylated analogue in the monolayer. A pI gradient composed of (NH(3)(+)/COO(-))-terminated thiols was also prepared and evaluated with respect to its ability to separate differently charged proteins, pepsin, and lysozyme, on a solid surface.

Microcontact printing (µCP) has been used to introduce temporary hydrophobic barriers on carboxymethylated dextran (CMD) hydrogels on gold. Among the investigated types of inks, tetraoctadecylammonium bromide (TOAB), electrostatically bound to the CMD layer, provided the most well-defined features both with respect to pattern-definition and reversibility upon exposure to a regeneration solution. The printed patterns were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), microscopic wetting and imaging null ellipsometry to explore the influence of concentration of ink solution and contact time on the appearance of the printed layer. AFM revealed that the printed TOAB molecules aggregated into clusters rather than into a homogeneous mono- or multilayer on the CMD hydrogel. It was also observed that printed areas of TOAB that are larger than 25?µm are inhomogeneous most likely because of an edge transfer lithography (ETL) mechanism. A protein model system based on Protein A-rabbit antimouse Fc ? was used to evaluate the potential of the patterned surface as a protein microarray chip by means of surface plasmon microscopy (SPM). Moreover, non-specific adsorption of several proteins onto TOAB barriers was also studied using surface plasmon resonance (SPR), and it is evident that undesired adsorption can be eliminated by removing barriers after ligand immobilization, but prior to analyte exposure, by treating the patterned surface with a simple salt regeneration solution. © Springer-Verlag/Wien 2004.

Two different types of ω-substituted alkanethiol/disulfide compounds have been used to prepare monolayer architectures on gold serving as platforms for the immobilization of receptor probe molecules – antibodies. These are: (i) carboxylic acid alkanethiols post-reacted with amino biotin to generate streptavidin surfaces, and (ii) N-hydroxysuccinimide-terminated disulfide surfaces. The properties of the monolayers, with and without attached receptor probe molecules, were analysed using infrared spectroscopy, ellipsometry, fluorescence scanning and atomic force microscopy. Several experimental parameters, such as condensation reagents, additives, probe and target concentrations and immobilization time, were systematically varied to determine the dynamic range and to optimize the sensitivity and signal-to-noise ratio of the biochip platforms. Fluorescence screening using Cy5-labelled antigens finally demonstrated that both surfaces could be successfully employed to immobilize the antibodies. The pros and cons of the two approaches are also discussed.

Temperature-programmed desorption (TPD) of self-assembled monolayers (SAMs) on gold is investigated by using in parallel mass spectrometry (MS) and infrared reflection−absorption spectroscopy (IRAS). Monolayers formed by HS(CH2)n−OH (n = 18, 22) and HS(CH2)15−CONH−(CH2CH2O)−H (EG1) are compared to reveal the influence of specifically introduced hydrogen-bonding groups on their thermal stability. The overall desorption process of the above molecules is found to occur in two main steps; a disordering of the alkyl chains followed by a complex series of decomposition/desorption reactions. The final step of the process involves desorption of sulfur from different chemisorption states. The amide-group-containing SAM, which is stabilized by lateral hydrogen bonds, displays a substantial delay of the alkyl chain disordering by about 50 K, as compared to the linear chain alcohols HS(CH2)n−OH. Moreover, the decomposition of the alkyls and the onset of sulfur desorption occur at a temperature that is higher by appro...

Localized surface plasmon resonance (LSPR) occurring in noble metal nanoparticles (e.g., Au) is a widely used phenomenon to report molecular interactions. Traditional LSPR sensors typically monitor shifts in the peak position or extinction in response to local refractive index changes in the close vicinity of the nanoparticle surface. The ability to resolve minute shifts/extinction changes is to a large extent limited by instrumental noise. A new strategy to evaluate LSPR responses utilizing changes in the shape of the extinction spectrum (the curvature) is proposed. The response of curvature to refractive index changes is investigated theoretically using Mie theory and an analytical expression relating the curvature to the refractive index is presented. The experimentally derived curvatures for 13 nm spherical gold nanoparticles (AuNPs) exposed to solvents with different bulk refractive indices confirm the theoretical predictions. Moreover, both the calculated and experimental findings suggest that the curvature is approximately a linear function of refractive index in regimes relevant to bio and chemical sensing. We demonstrate that curvature is superior over peak shift and extinction both in terms of signal-to-noise (S/N) ratio and reliability of LSPR sensors. With a curvature, one could readily monitor submonolayer adsorption of a low molecular weight thiol molecule (M(w) = 458.6) onto 13 nm AuNPs. It is also worthwhile mentioning that curvature is virtually insensitive to instrumental instabilities and artifacts occurring during measurement. Instabilities such as baseline tilt and shift, shift in peak position as well as sharp spikes/steps in the extinction spectra do not induce artifacts in the sensorgrams of curvature.

An approach based on electrochemistry to differentiate between phosphorylated and nonphosphorylated amino acid analogues adsorbed on gold is presented. Analogues of serine, threonine, and tyrosine, containing thiohexadecyl headgroups, were synthesized and assembled on gold, and the surface capacitance was evaluated using electrochemical impedance spectroscopy. A procedure for deprotection of tert-butyl phosphate protecting groups, on the monolayer, is also described. Characterizations of the assembled analogues by cyclic voltammetry, infrared spectroscopy, and ellipsometry are used to confirm the insulating properties of the monolayers and the outcome of surface modifications. The results from cyclic voltammetry show good insulating properties for the monolayers even after phosphate deprotection. The infrared measurements reveal well-ordered monolayers, and the thickness from ellipsometry is in good agreement with expectations from molecular modeling. The impedance experiments show a capacitance increase up to 0.6 microF/cm2 as phosphate groups are introduced. The results in this study indicate the possibility of using a surface chemical and impedance spectroscopy approach to detect the kinase/phosphatase activity and kinetics involved in phosphorylation reactions.

The coagulation of blood plasma and whole blood was studied with a surface plasmon resonance (SPR) based device and a quartz crystal microbalance instrument with energy dissipation detection (QCM-D). The SPR and QCM-D response signals were similar in shape but differing in time scales, reflecting differences in detection mechanisms. The QCM-D response time was longer than SPR, as a physical coupling of the sample to the substrate is required for molecules to be detected by the QCM-method. Change of sample properties within the evanescent field is sufficient for detection with SPR. Both the SPR signals and the QCM-D frequency and dissipation shifts showed dependency on concentrations of coagulation activator and sensitivity to heparin additions. The ratio of dissipation to frequency shifts, commonly considered to reflect viscoelastic properties of the sample, varied with the concentration of activator in blood plasma but not in whole blood. Additions of heparin to the thromboplastin activated whole blood sample, however, made the ratio variation reoccur. Implications of these observations for the understanding of the blood coagulation processes as well as the potential of the two methods in the clinic and in research are discussed.

This paper explores the interfacial properties of one-dimensional molecular gradients of alkanethiols (HS‐(CH2)n ‐ X) on gold. The kinetics and thermodynamics of monolayer formation are important issues for these types of mixed molecular assemblies. The influence of chain length difference on the contact angles with hexadecane (HD), ua and ur, and the hysteresis, has been studied by employing alkanethiols HS‐(CH2)n‐CH3, with n 9, 11, 13, 15 and 17, in the preparation of the self-assembled monolayers (SAM) gradients. The contact angles with hexadecane, at the very extreme ends of the gradients, show characteristic values of a highly ordered CH3-like assembly: ua 45‐50°. In the middle of the gradients ua drops noticeably and exhibits values representative for CH2-like polymethylenes, ua20‐30°, indicating a substantial disordering of the protruding chains of the longer component in the gradient assembly. As expected, the exposure of CH2-groups to the probing liquid increases with increasing differential chain length of the two n-alkanethiol used, in this case eight methylene units. However, the contact angles always display a non-zero value which means that even at a chain length difference of eight methylene units there is a substantial exposure of methyl (CH3) groups to the probing liquid. With infrared reflection-absorption spectroscopy (IRAS) we have monitored the structural behavior of the polymethylene chains along the gradient. We find complementary evidence for disordered chains in the gradient region, and the IRAS results correlate well with the contact angle measurements. © 1999 Elsevier Science B.V. All rights reserved.

Humidity influences the adsorption of dimethyl methylphosphonate (DMMP) on three different organic model interfaces to quite different extents. The model interfaces, characterized by the following tail groups (-OH CH 3 , -COOH), were prepared by solution self-assembly of w-substituted alkanethiols on gold, and the coadsorption of water and DMMP was followed by a surface acoustic wave sensor and infrared reflection absorption spectroscopy. The surfaces show both quantitative and qualitative differences concerning the interaction with DMMP. Humidity is found to influence the hydrogen bonds formed between DMMP and the tail groups, increasing the strength ofinteraction. However, the changes in surface coverage of DMMP, as measured by infrared absorption, are not fully registered by the surface acoustic wave sensor, because of a concomitant desorption of water. Indeed there is even a loss in sensor signal due to the presence of humidity in the case of the -COOH-terminated surface, where the strongest interaction with DMMP is found. After compensation for the loss of water, the coverage of DMMP increases with the relative humidity on the OH surface and decreases with the relative humidity on the COOH surface. The consequences of the results are discussed in terms of their impact on the design of sensing interfaces based on hydrogen bonding.

The adsorption of 3-(2-thioethyl)pyrrole (3-TEP) and 1-(2-thioethyl)pyrrole (1-TEP) on gold has been studied in ultrahigh vacuum with infrared reflection absorption spectroscopy, X-ray photoelectron spectroscopy, and temperature-programmed desorption mass

Surfaces prepared by self-assembly of alkanethiolates onto thin (10 nm) gold films supported on glass have been used as substrates for surface force measurements between macroscopic surfaces. Surface roughness, the order in the monomolecular film, wetting properties, and their stability in aqueous electrolyte solutions have been investigated using atomic force microscopy, infrared absorption spectroscopy, and contact angle measurements. Direct force measurements have been performed with a noninterferometric bimorph surface force apparatus, using surfaces with differently functionalized thiolates; measurements were made in air, water, and aqueous electrolyte solutions. Results from force measurements between hydrophobic and neutral hydrophilic surfaces in air and water are presented and discussed, as well as some results from measurements with surfaces exposing carboxylic groups in various aqueous electrolyte solutions. It is demonstrated that alkanethiol monolayers self-assembled onto thin gold films on m...

The development of accurate analytical tools to control the interfacial properties of solid substrates is of importance for the design of new biomaterials, as well as for the understanding of biomolecular interactions on surfaces. Considerable research efforts are presently devoted to this area on different levels of molecular complexity, i.e. both in the presence and in the absence of the biomolecules. In this contribution we review briefly applications of infrared reflection–absorption spectroscopy (IRAS) and ellipsometry as tools for analysis of the chemical properties of model surfaces, and their biological response in vitro when in contact with blood plasma or serum, respectively. The strength of the combination of the techniques is demonstrated by determination of pro-tein adsorption patterns on a series of chemically well-defined so-called self-assembled alkanethiolate monolayers (SAMs) of 16-thiohexadecanol (HS–(CH 2 ) 16 –OH) and n -hexadecanethiol (HS–(CH 2 ) 15 –CH 3 ) on gold. The protein adsorption patterns after incubations in plasma were determined by the specific binding of antibodies to the surfaces.

Botulinum neurotoxin is considered as one of the most toxic food-borne substances and is a potential bioweapon accessible to terrorists. The development of an accurate, convenient, and rapid assay for botulinum neurotoxins is therefore highly desirable for addressing biosafety concerns. Herein, novel biotinylated peptide substrates designed to mimic synaptosomal-associated protein 25 (SNAP-25) are utilized in gold nanoparticle-based assays for colorimetric detection of botulinum neurotoxin serotype A light chain (BoLcA). In these proteolytic assays, biotinylated peptides serve as triggers for the aggregation of gold nanoparticles, while the cleavage of these peptides by BoLcA prevents nanoparticle aggregation. Two different assay strategies are described, demonstrating limits of detection ranging from 5 to 0.1 nM of BoLcA with an overall assay time of 4 h. These hybrid enzyme-responsive nanomaterials provide rapid and sensitive detection for one of the most toxic substances known to man. ASTAR (Agency for Sci., Tech. and Research, S’pore) Accepted version

We report herein on the employment of synthetic peptide-functionalized gold nanoparticles (AuNPs) with various diameters as radiative quenchers for the time-resolved monitoring of botulinum A light chain (BoLcA) activity. The results demonstrate that larger AuNPs provide higher energy transfer efficiencies between the dye and the AuNPs, but poorer BoLcA activities for the proteolysis of peptides because of steric constraints. The initial turnover number for the BoLcA proteolysis of peptides on 18 nm AuNPs was retarded by a factor of 80 as compared with 1.4 nm AuNPs. A similar phenomenon has been observed for trypsin, however, with less hindrance on large AuNPs. Thus, the use of smaller 1.4 nm AuNPs in conjunction with robust synthetic peptides provides an attractive format for the time-resolved monitoring of protease activity and for BoLcA sensing at a highly competitive limit of detection (1 pM). ASTAR (Agency for Sci., Tech. and Research, S’pore) Accepted version

Molecular gradients of ω-substituted alkanethiols HS(CH 2 ) 15 X (X:CH 3 , CH 2 OH, CO 2 cH 3 , CH 2 C=N) were prepared on sputtered gold films by using a novel cross-diffusion methodology. Ellipsometry, contact angle measurements, infrared spectroscopy, and X-ray photoelectron spectroscopy (XPS) were used to monitor the chain conformation and gradient profiles. The XPS analysis were with improved lateral resolution as compared to our earlier infrared study (Langmuir 199o-, 11, 3821) by imaging a 160 μm wide segment of the gradient surface on the slit of the XPS analyzer and by using a step size of 0.5 mm. The so-obtained gradient profiles are expected to be more representative of the true composition profiles and were for the three gradients investigated found to be 5-6 mm, about 2-3 shorter than the corresponding profiles obtained by scanning infrared spectroscopy. The coverages alkyl chain densities) were found to be constant for the gradients prepared from thiols with relatively small-tail groups C=N and CH 3 , whereas a minor reduction in coverage, 3-4%, was observed in a narrow range for the gradient containing the CO 2 CH 3 -terminated alkanethiol. This change is, however, too small to be by ellipsometry and infrared spectroscopy and supports a model where the gradient assemblies approach the limiting structure of densely packed all trans polymethylenes.

Four different carbonyl-containing self-assembled monolayers (SAMs) of alkanethiolates on gold were studied to assess the impact of the functional group linked to the carbonyl upon its hydrogen bond accepting capability. These SAMs (HS(CH 2 ) 16 O(C=O)-X, X = CH 3 , CF 3 , or C 6 H 5 , and HS(CH 2 ) 15 (C=O)OCH 3 ) were thoroughly characterized with contact angle measurements, single wavelength ellipsometry, and infrared reflection-absorption spectroscopy (IRAS) prior to the studies of interaction with D 2 O. The first three monolayer compounds were introduced by reacting hydroxyl-terminated SAMs (HS(CH 2 ) 16 OH with either acetyl chloride, trifluoroacetic anhydride, or benzoyl chloride. The behavior of D 2 O ice on the SAMs was investigated at 100 K with IRAS and temperature programmed desorption (TPD). On all monolayers the D 2 O molecules were shown to interact with the carbonyl oxygen. The degree of interaction depended upon the termination of the thiol, where the size, structure, and electronegativity of the terminating groups in the molecules comprising the monolayer were found to be important factors. Indications of interaction with the C-O-C oxygen were seen for all compounds, as well as weak interaction between water molecules and the CF 3 group of one of the investigated SAMs. Common behavior for all four monolayers with an adsorbed D 2 O overlayer was a decrease in the number of hydrogen bonds to the substrate when the overlayer was annealed from amorphous ice at 100 K to polycrystalline-like ice at 140 K. The spectral changes accompanying the structural transition were consistent with a change from a mainly flat overlayer to condensed three-dimensional clusters. The bulk-to-surface molecular ratio of adsorbed ice clusters could be assessed by IRAS and correlated to macroscopic wetting properties. Our results infer that microscopic ice clusters on these SAMs qualitatively mimic the shape of macroscopic water drops on the same SAMs. Results of TPD measurements are also consistent with this view.

A self-assembled monolayer (SAM) on gold prepared from a binary mixture of a thiol analogue of cholesterol (thiocholesterol, TC) and a functionalised alkanethiol (11-mercaptoundecanoic acid, MUA) has been investigated by voltammetry. The voltammetric results are in agreement with previously reported spectroscopic data and show that the geometric arrangement and composition of the molecules in the mixed monolayer controls the heterogeneous electron transfer process of Fe(CN)63− across the assembly. The quantitative description of the influence of TC on the electron transfer rate constant is given through Tafel plots. At the oure MUA SAM electrode, the electron transfer is governed by penetration through the monolayer. The introduction of TC into the SAMs creates defects giving rise to diffusion controlled electron transfer in addition to penetration. By raising the TC content the electron transfer rate constant increases due to diffusion. This behaviour can be explained by a model in which the assembly goes from a penetrative but defect-free film barrier (pure MUA SAM) via a structure with defects in the mixed composition regime to a defect-rich structure consisting of an array of ultramicroelectrodes (pure TC SAM).

Infrared reflection–absorption spectroscopy has been used to characterize thin overlayers (1–200 A) of D2O ice deposited in UHV onto a set of self-assembled alkanethiolate monolayers (SAMs) of controlled wettabilities on gold. The SAMs were prepared from a series of controlled composition, mixed solutions of HS(CH2)15CH3 and HS(CH2)16OH, making it possible to investigate the whole wettability range from θ≈0° to θ=112°, where θ is the static contact angle with water. Dosing of D2O and infrared measurements were carried out at selected sample temperatures between 82 and 150 K. Experimental spectra of ice overlayers recorded below 100 K on all SAM substrates are in good agreement with simulated reflection–absorption spectra, derived from the optical constants of amorphous ice. This agreement allows accurate film thickness determination. In contrast, lack of correspondence in spectral signature is noted between the spectra of annealed films and simulated polycrystalline (or amorphous) ice spectra. We interpre...

Infrared reflection−absorption spectroscopy is used to investigate thin (1−200 A average thickness) overlayers of D2O ice deposited in ultrahigh vacuum on controlled wettability self-assembled monolayers. The monolayers were derived from mixed solutions of HS(CH2)15CH3 and HS(CH2)16OH, making it possible to examine the whole range of samples from fOH = 0.0 to fOH = 1.0, where fOH denotes the molar fraction of OH-terminated thiols in the preparation solution. This paper focuses on the interaction between the ice and the monolayer. It is shown that water molecules do not penetrate into the monolayer but that two types of interaction with the chain-terminating groups occur: hydrogen bond formation with surface hydroxyls and weak dipole−dipole interaction with surface methyls. For surfaces with fOH < 0.3, the latter interaction causes the free OD mode, normally observed at 2729 cm-1, to shift to 2704 cm-1, thereby providing a spectral signature feature whose intensity is directly proportional to the relative...

The formation of thiocholesterol (TC) monolayers on gold has been studied by ellipsometry, contact angle measurements, infrared spectroscopy, and cyclic voltammetry. Subsequent treatment of the TC assembly with 11-mercaptodeuterioundecanoic acid (MDUA) shows that the average surface coverage is about 65% of that of a self-assembled alkanethiolate monolayer and that it has a large number of molecular defects. These defects exist because of a mismatch between the size and shape of the TC molecule and the pinning distance at the Au(111) crystal lattice. Potential uses of these defect-rich structures are microelectrode arrays for electroanalytical and biosensor applications.

Equimolar, binary-component, self-assembled monolayers (SAMs) of -S(CH 2 ) 15 CH 2 OH and -S(CH 2 ) (15+m) -CH 3 , with m varied systematically from -6 to +6, were prepared on evaporated gold films by self-assembly from ethanol solutions of the corresponding thiols. Single-wavelength ellipsometry, X-ray photoelectron spectroscopy, and infrared reflection spectroscopy (IRS) were used to monitor the coverage, composition, and chain conformational order, respectively, in the air/SAM structures, while liquid drop contact angle measurements with hydrogen-bonding and hydrocarbon liquids were used to probe the structural and chemical characteristics at the liquid/SAM interface. Both the IRS C-H stretching mode characteristics and the hydrocarbon liquid contact angles show sharp changes as m increases through the region of -4 to 0, while the hydrogen-bonding liquid contact angles remain essentially constant across the range of m values. The combined data lead to a general structural model in which monolayer structures shift from randomly placed, protruding HO-terminated chain segments which screen the underlying CH 3 surface at low m to contiguously placed, protruding CH 3 -terminated chains, organized in clusters and extending away from the underlying CH 2 OH surface at high m. These structures require that the monolayers approach random mixing at low m but be phase-segregated at high m.

A general methodology capable of generating a molecular, chemical concentration gradient on a solid substrate in one surface dimension is presented. Although the method can be extended to more than one surface dimension, and to a variety of molecule/substrate systems, we have chosen to demonstrate it by utilizing thiols on gold as the model system. The gradients are prepared by cross-diffusing ω-substituted alkanethiols, HS(CH 2 ) n X (X = OH, CO 2 CH 3 , COOH, and CD 3 , n = 10-19), with or without fully deuterated chains, and simple n-alkanethiols, HS(CH 2 ) m Y (Y = CH 3 , m = 9-17), toward each other from opposite ends of a polysaccharide matrix deposited on top of a gold substrate. The so-prepared gradients are characterized by a number of techniques, including ellipsometry, contact angle measurements, and infrared reflection absorption spectroscopy (IRAS). The observed ellipsometric thicknesses and the limiting contact angles of the gradient assemblies agree very well with the values obtained from the corresponding single component and mixed self-assembled monolayers. The infrared data furthermore suggest that the entire gradient is composed ofwell-organized and densely packed all-trans hydrocarbon chains for thiols with chain length >10. The lateral dimensions of the gradient regions vary between 4 and 8 mm for the combination diffusion pairs employed in this study. The upper and lower values account for diffusion pairs with long and short hydrocarbon chains, respectively. It must be stressed, however, that the infrared and ellipsometric techniques only can provide information about the molecular organization on the macroscopic scale because of the relatively large spot size (1 mm) used in the experiments. Scanning probe microscopy must be employed to further investigate the mechanism of gradient formation, the molecular concentration profile, and mixing behavior on the microscopic (molecular) scale. We believe, however, that the large number of chemical and structural combinations attainable with the present approach can be used to address a broad range of surface physics/chemistry problems in material science and generate novel and unique model organic architectures. We discuss briefly the potential applications ofthese novel gradients in areas of relevance to molecular recognition and biosensing.

This paper examines the relationship between the thermal desorption of thin overlayers of condensed D2O ice and the wettability properties of the supporting substrate surface. Mixed self-assembled ...

Tricyclohexylphosphine adsorbates on gold, as well as multilayer films of tricyclohexylphosphine, are investigated by X-ray photoelectron spectroscopy (XPS) and infrared reflection-absorption spectroscopy (IRAS). Molecular orientation and molecular surface interaction for adsorbates prepared from solution are studied, using the multilayer films as references. Since phosphines are easily oxidized in contact with air, the reference, made from a pressed pellet of the starting material, consists of a considerable amount of oxidized tricyclohexylphosphine. Therefore an oxygen-free reference would also be desirable. Such a reference of a tricyclohexylphosphine multilayer was obtained by fractionating tricyclohexylphosphine from tricyclohexylphosphine oxide by a sublimation process under ultrahigh vacuum. The results from both a pressed pellet of the starting material and an evaporated tricyclohexylphosphine multilayer are compared with the results from the adsorbate. When tricyclohexylphosphine is adsorbed from solution onto gold, there is a preferential adsorption of the unoxidized tricyclohexylphosphine. The molecular structure is intact during adsorption and the molecules are oriented with the phosphine atoms close to the surface. The chemical shift of the P(2p 3/2 ) binding energy during adsorption indicates an electron donation from the adsorbate to the meta