Thiol-ene micropillar array electrospray ionization platform for zeptomole level bioanalysis.

A micropillar array electrospray ionization (µPESI) platform fabricated from thiol-enes with 56 individual polyethylene glycol coated µPESI chips for bioanalytical mass spectrometry is introduced. Bioanalysis capability is shown by measurement of a protein, a protein digest and a cell lysate sample. The thiol-ene polyethylene glycol (PEG) coated µPESI chip allows the use of a wide range of aqueous-organic solvent compositions and provides a detection limit at 60 zeptomole level (6 × 10-20 mol) for a peptide standard.

Robust hybrid elastomer/metal-oxide superhydrophobic surfaces.

We introduce a new type of hybrid material: a nanostructured elastomer covered by a hard photoactive metal-oxide thin film resembling the exoskeleton of insects. It has extreme water repellency and fast self-recovery after damage. A new fabrication method for replicating high aspect ratio, hierarchical re-entrant aluminum structures into polydimethylsiloxane (PDMS) is presented. The method is based on a protective titania layer deposited by atomic layer deposition (ALD) on the aluminum template. The ALD titania transfers to the elastomeric scaffold via sacrificial release etching. The sacrificial release method allows for high aspect ratio, even 100 µm deep and successful release of overhanging structures, unlike conventional peeling. The ALD titania conformally covers the 3D multihierarchical structures of the template and protects the polymer during the release etch. Afterwards it prevents the high aspect ratio nanostructures from elasticity based collapse. The resulting nanostructured hybrid PDMS/titania replicas display robust superhydrophobicity without any further fluoro-coating or modification. Their mechanical and thermal robustness results from a thick nanostructured elastomeric layer which is conformally covered by ceramic titania instead of a monolayer hydrophobic coating. We have demonstrated the durability of these replicas against mechanical abrasion, knife scratches, rubbing, bending, peel tape test, high temperature annealing, UV exposure, water jet impingement and long term underwater storage. Though the material loses its superhydrophobicity in oxygen plasma exposure, a fast recovery from superhydrophilic to superhydrophobic can be achieved after 20 min UV irradiation. UV-assisted recovery is correlated with the high photoactivity of ALD titania film. This novel hybrid material will be applicable to the large area superhydrophobic surfaces in practical outdoor applications.

Stroboscopic supercontinuum white-light interferometer for MEMS characterization.

We used a supercontinuum-based scanning white-light interferometer to characterize the oscillation of a MEMS device. The output of a commercially available supercontinuum light source (FiberWare Ilum II USB) was modulated to achieve stroboscopic operation. By synchronizing the modulation frequency of the source to the sample oscillation, dynamic 3-D profile measurements were recorded. These results were validated against those obtained with a white light LED setup. The measured maximum deflection of a 400×25×4 µm(3) microbridge driven with 0-6.8 V sinusoidal voltage at 10 Hz was 1.42±0.03 µm (supercontinuum), which agreed with the LED measurement. The method shows promise for characterization of high-frequency MEMS devices.

Immunoassay of C-reactive protein by hot electron induced electrochemiluminescence using integrated electrodes with hydrophobic sample confinement.

C-reactive protein (CRP) was determined in the concentration range 0.01-10 mg L(-1) using hot electron induced electrochemiluminescence (HECL) with devices combining both working and counter electrodes and sample confinement on a single chip. The sample area on the electrodes was defined by a hydrophobic ring, which enabled dispensing the reagents and the analyte directly on the electrode. Immunoassay of CRP by HECL using integrated electrodes is a good candidate for a high-sensitivity point-of-care CRP-test, because the concentration range is suitable, miniaturisation of the measurement system has been demonstrated and the assay method with integrated electrodes is easy to use. High-sensitivity CRP tests can be used to monitor the current state of cardiovascular disease and also to predict future cardiovascular problems in apparently healthy people.

UV-embossed inorganic-organic hybrid nanopillars for bioapplications.

We demonstrate rapid and maskless wafer-scale fabrication of inorganic-organic hybrid nanopillars for microfluidic and mass spectrometric applications. The nanopillar features are transferred from a black silicon master to the hybrid material by UV-embossing with a composite stamp made of polydimethylsiloxane (PDMS)/hard-PDMS (h-PDMS), which allows accurate transfer of structures in the 100 nm size range. The nanopillar embossing process is extremely simple and well suited for cheap wafer-scale prototyping, as the process does not require expensive masks or stamps and even works with a standard office lamp as the exposure source. The pillars can also be fabricated on a wide variety of substrates because the hybrid material is solvent-free. The use of the hybrid nanopillars as a hydrophobic/philic platforms and as sample plates in surface assisted laser desorption ionization mass spectrometry is demonstrated.

Coating of nanoporous membranes: atomic layer deposition versus sputtering.

Nanoporous anodic alumina membranes and silicon samples with plasma etched nanopores have been coated with zinc oxide or gold layer using atomic layer deposition (ALD) or sputtering, respectively. In the case of ALD process, the precursor pulses were extended, compared with planar substrate coating. Thick (60 microm) anodic alumina membranes have been conformally coated with zinc oxide ALD layer. Metal sputtering technique was used just for opposite purpose--to minimize the penetration of gold into the pores during gold-coating of the top and bottom surfaces of the membrane. Scanning electron microscopy (SEM) has been used to investigate the layer thickness, uniformity and conformality inside the nanopores.

The fabrication of silicon nanostructures by local gallium implantation and cryogenic deep reactive ion etching.

We show that gallium-ion-implanted silicon serves as an etch mask for fabrication of high aspect ratio nanostructures by cryogenic plasma etching (deep reactive ion etching). The speed of focused ion beam (FIB) patterning is greatly enhanced by the fact that only a thin approx. 30 nm surface layer needs to be modified to create a mask for the etching step. Etch selectivity between gallium-doped and undoped material is at least 1000:1, greatly decreasing the mask erosion problems. The resolution of the combined FIB-DRIE process is 20 lines microm(-1) with the smallest masked feature size of 40 nm. The maximum achieved aspect ratio is 15:1 (e.g. 600 nm high pillars 40 nm in diameter).

Feasibility of atmospheric pressure desorption/ionization on silicon mass spectrometry in analysis of drugs.

The feasibility of atmospheric pressure desorption/ionization on silicon mass spectrometry (AP-DIOS-MS) for drug analysis was investigated. It was observed that only compounds with relative high proton affinity are efficiently ionized under AP-DIOS conditions. The limits of detection (LODs) achieved in MS mode with midazolam, propranolol, and angiotensin II were 80 fmol, 20 pmol, and 1 pmol, respectively. In MS/MS mode the LODs for midazolam and propranolol were 10 fmol and 5 pmol, respectively. The good linearity (r(2) > 0.991), linear dynamic range of 3 orders of magnitude, and reasonable repeatability showed that the method is suitable for quantitative analysis.

Poly(dimethylsiloxane) electrospray devices fabricated with diamond-like carbon-poly(dimethylsiloxane) coated SU-8 masters.

This study presents coupling of a poly(dimethylsiloxane) (PDMS) micro-chip with electrospray ionization-mass spectrometry (ESI-MS). Stable electrospray is generated directly from a PDMS micro-channel without pressure assistance. Hydrophobic PDMS aids the formation of a small Taylor cone in the ESI process and facilitates straightforward and low-cost batch production of the ESI-MS chips. PDMS chips were replicated with masters fabricated from SU-8 negative photoresist. A novel coating, an amorphous diamond-like carbon-poly(dimethylsiloxane) hybrid, deposited on the masters by the filtered pulsed plasma arc discharge technique, improved significantly the lifetime of the masters in PDMS replications. PDMS chip fabrication conditions were observed to affect the amount of background peaks in the MS spectra. With an optimized fabrication process (PDMS curing agent/silicone elastomer base ratio of 1/8 (w/w), curing at 70 degree C for 48 h) low background spectra were recorded for the analytes. The performance of PDMS devices was examined in the ESI-MS analysis of some pharmaceutical compounds and amino acids.

Preparation of porous n-type silicon sample plates for desorption/ionization on silicon mass spectrometry (DIOS-MS).

This study focuses on porous silicon (pSi) fabrication methods and properties for desorption ionization on silicon mass spectrometry (DIOS-MS). PSi was prepared using electrochemical etching of n-type silicon in HF-ethanol solution. Porous areas were defined by a double-sided illumination arrangement: front-side porous areas were masked by a stencil mask, eliminating the need for standard photolithography, and backside illumination was used for the backside ohmic contact. Backside illumination improved the uniformity of the porosified areas. Porosification conditions, surface derivatizations and storage conditions were explored to optimize pSi area, pore size and pore depth. Chemical derivatization of the pSi surfaces improved the DIOS-MS performance providing better ionization efficiency and signal stability with lower laser energy. Droplet spreading and drying patterns on pSi were also examined. Pore sizes of 50-200 nm were found to be optimal for droplet evaporation and pore filling with the sample liquid, as measured by DIOS efficiency. With DIOS, significantly better detection sensitivity was obtained (e.g. 150 fmol for midazolam) than with desorption ionization from a standard MALDI steel plate without matrix addition (30 pmol for midazolam). Also the noise that disturbs the detection of low-molecular weight compounds at m/z < 500 with MALDI could be clearly reduced with DIOS. Low background MS spectra and good detection sensitivity at the 100-150 fmol level for pharmaceutical compounds were achieved with DIOS-MS.

Hospital mouth-cleaning aids may cause dental erosion.

Maintaining satisfactory oral health in bed-ridden patients often calls for special aids, such as moistened cotton swabs. The purpose of this in vitro study was to investigate three commercially available swabsticks meant for hospital use (Dentiswab, Lemon-Glycerin Swabsticks, and Lemon Glycerine Swabs), and one saliva-stimulant chewing tablet (Cassisal), regarding their possible erosive effects on dental enamel. A bovine enamel model system was used. Specimens were incubated for 4 hours in the test solutions made of the products. The results showed statistically significant enamel softening caused by two of the cotton swabs (Lemon-Glycerin Swabsticks and Lemon Glycerine Swabs). Incubation in the solutions made of Dentiswab cotton swabs and Cassisal tablets caused only minimal changes in microhardness. Profilometric measurements supported these results. Stereomicroscopy and SEM showed distinct erosion in specimens treated with the lemon swab solution, while the other two did not cause erosion. Thus, attention should be focused on choosing the less erosive products if special mouth-cleaning aids are used.

Oral hygiene products may cause dental erosion.

The erosive effects of the mouthwashes Calcusan, Salisan and Veadent, and of a tablet, Salivin, meant to be sucked, were studied using enamel blocks from bovine permanent teeth. For comparison, the Hart-Sport sport drink (pH 3.1), which is known to be erosive, and carbonated mineral water Vichy (pH 5.4), which is known to have no erosive effect, were also tested. After two hours of exposure to each test product losses of tooth material were measured by recording the surface profile, using a Dectak profilometer. Changes in the enamel blocks were also studied using a stereomicroscope and a scanning electron microscope (SEM). In the profilometric study, Calcusan was found to have a marked erosive effect (77 +/- 5 microns), four times as severe as the erosive effect of the positive control Hart-Sport (19 +/- 4 microns). Veadent had a less marked effect (3 +/- 1 microns). Salisan and Salivin had no detectable erosive effects and were similar in action to the negative control, carbonated mineral water. From the SEM pictures, Calcusan resulted in an EDTA-type reaction, with dissolution of the peripheral areas of enamel prisms. It is important to recognize that the organic acids and chelating agents in various oral hygiene products may have deleterious effects on teeth.