Chemistries for Making Additive Nanolithography in OrmoComp Permissive for Cell Adhesion and Growth.

Two-photon lithography allows writing of arbitrary nanoarchitectures in photopolymers. This design flexibility opens almost limitless possibilities for biological studies, but the acrylate-based polymers frequently used do not allow for adhesion and growth of some types of cells. Indeed, we found that lithographically defined structures made from OrmoComp do not support E18 murine cortical neurons. We reacted OrmoComp structures with several diamines, thereby rendering the surfaces directly permissive for neuron attachment and growth by presenting a surface coating similar to the traditional cell biology coating achieved with poly-d-lysine (PDL) and laminin. However, in contrast to PDL-laminin coatings that cover the entire surface, the amine-terminated OrmoComp structures are orthogonally modified in deference to the surrounding glass or plastic substrate, adding yet another design element for advanced biological studies.

Ethnic hair care products may increase false positives in hair drug testing.

The question of why different races appear more susceptible to hair contamination by external drugs remains controversial. This research studied susceptibility of head hair to external cocaine and methamphetamine when hair products have been applied. Three different chemical classes of ethnic hair products were applied to Caucasian, Asian, and African hair. Some products increased the methamphetamine and cocaine concentrations in all hair types. A unique finding of this research is that certain ethnic hair products can replace moisture as a diffusion medium, thereby increasing the susceptibility to contamination over 100-fold compared to petroleum-based products.

Functionalization of micro- and nano-apertures with chromate-selective solvent polymeric membrane.

A new miniaturization approach to create micro- and nanoscale ion selective electrodes (ISEs) was demonstrated and the concept tested with an environmentally relevant chromate-selective membrane consisting of 7.7:62.2:31.1 wt % Aliquat336:2-NPOE:PVC. Apertures of 100 nM and 30 microM dimensions were made using MEMS fabrication techniques and functionalized through a macroscale application of solvent polymeric membrane. Performance studies for the microscale ISE showed a response slope of -58.6+/-5.6 mV decade(-1) and limit of detection (LOD) of 2.1 x 10(-5)+/-1.1 x 10(-5) M, versus -65.2+/-4.2 mV decade(-1) and 1.8 x 10(-5)+/-6 x 10(-6) M for the nanoscale ISE. This was consistent with control studies with carefully conditioned coated wire electrodes, which demonstrated a response slope of -61.7+/-2.4 mV decade(-1) and a LOD of 3.0 x 10(-6)+/-1 x 10(-6) M. Response times for the best micro- and nanoscale ISEs were in the 10-20 s timeframe. Electrical resistance measurements were in the GOmega range for the microscale ISEs and nanoscale ISEs. Appropriate ISE geometry was confirmed through AFM measurements and calculations based on electrical properties for micro- and nanoscale apertures. These micro- and nanoscale ISEs are expected to have significant impact in the field of microscale analytical processes.

Bilayer lipid membrane (BLM) based ion selective electrodes at the meso-, micro-, and nano-scales.

This paper presents a novel method for making micron-sized apertures with tapered sidewalls and nano-sized apertures. Their use in bilayer lipid membrane-based ion selective electrode design is demonstrated and compared to mesoscale bilayers and traditional PVC ion selective electrodes. Micron-sized apertures are fabricated in SU-8 photoresist films and vary in diameter from 10 to 40 microm. The tapered edges in SU-8 films are desired to enhance bilayer lipid membrane (BLM) formation and are fabricated by UV-light overexposure. Nano-apertures are made in boron diffused silicon film. The membranes are used as septa to separate two potassium chloride solutions of different concentrations. Lecithin BLMs are assembled on the apertures by ejecting lipid solution. Potassium ionophore, dibenzo-18-crown-6, is incorporated into BLMs by dissolving it in the lipid solution before membrane assembly. Voltage changes with increasing potassium ion concentrations are recorded with an A/D converter. Various ionophore concentrations in BLMs are investigated. At least a 1% concentration is needed for consistent slopes. Electrode response curves are linear over the 10(-6) to 0.1M range with a sub-Nernstian slope of 20mV per Log concentration change. This system shows high selectivity to potassium ions over potential interfering sodium ions. BLMs on the three different aperture sizes at the meso-, micro-, and nano-scales all show similar linear ranges and limits of detection (LODs) as PVC ion selective membranes.

Fourier transform infrared spectroscopy of azide and cyanate ion pairs in AOT reverse micelles.

Evidence for ion pair formation in aqueous bis(2-ethylhexyl) sulfosuccinate (AOT) reverse micelles (RMs) was obtained from infrared spectra of azide and cyanate with Li(+), Na(+), K(+), and NH(4)(+) counterions. The anions' antisymmetric stretching bands near 2000 cm(-1) are shifted to higher frequency (blueshifted) in LiAOT and to a lesser extent in NaAOT, but they are very similar to those in bulk water with K(+) and NH(4)(+) as the counterions. The shifts are largest for low values of w(o) = [water]/[AOT] and approach the bulk value with increasing w(o). The blueshifts are attributed to ion pairing between the anions and the counterions. This interpretation is reinforced by the similar trend (Li(+)>Na(+)>K(+)) for producing contact ion pairs with the metal cations in bulk dimethyl sulfoxide (DMSO) solutions. We find no evidence of ion pairs being formed in NH(4)AOT RMs, whereas ammonium does form ion pairs with azide and cyanate in bulk DMSO. Studies are also reported for the anions in formamide-containing AOT RMs, in which blueshifts and ion pair formation are observed more than in the aqueous RMs. Ion pairs are preferentially formed in confined RM systems, consistent with the well established ideas that RMs exhibit reduced polarity and a disrupted hydrogen bonding network compared to bulk water and that ion-specific effects are involved in mediating the structure of species at interfaces.

Measuring quaternary ammonium cleaning agents with ion selective electrodes.

Data for coated-wire, ion selective electrodes (ISEs) are presented for cationic surfactant ions found in common cleaners including benzyldimethyltetradecylammonium, benzyldimethyldodecylammonium, and benzyldimethylhexadecylammonium. The ion exchangers dinonylnaphthalene sulfonic acid, tetraphenyborate, and tetrakis(4-chlorophenyl)borate are examined, showing dinonylnaphthalene sulfonic acid to be the favored species. The ISEs exhibit approximately Nernstian behavior down to the 10(-6) M limit of detection with lifetimes in excess of 50 days when used continuously, and a shelf life of over 100 days. Reaching the upper detection limit at the critical micelle concentration requires use of polymeric-membrane reference electrodes including a new membrane cocktail, which allow response measurements of an order of magnitude higher than the traditional fritted-glass reference electrode. The surfactant ISEs show excellent selectivity over the common metal ions Na+, K+, Mg2+, Ca2+, and Cu2+ with selectivity coefficients less than 10(-5.3). The ISEs are also selective over the lower molecular weight quaternary ammonium ions tetradecyltrimethylammonium, dodecyltrimethylammonium, benzyldimethyl(2-hydroxyethyl)ammonium, and tetrabutylammonium with selectivity coefficients ranging from 10(-1.7) to 10(-5.5). Use of a single electrode to determine accurately the total cationic surfactant concentration in common cleaning solutions is accomplished with information about concentration dependent interferences and a modified Nikolsky-Eisenman model. Finally, quaternary ammonium surfactants have a deleterious effect on the measurements of pH and common ions like K+, Mg2+ and Ca2+ with polymeric ISEs. This makes it critical to include surfactant electrodes in a detector array when cleaning agents are present.

An enzyme electrode for amperometric measurement of D-amino acid.

A carbon paste enzyme electrode has been developed for measurement of D-amino acids that employs a fatty acid modified FAD to prevent leaching of this essential cofactor to the surrounding aqueous environment and which serves as an enzyme stabilizing agent. The lower limit of detection is at least 10(-4) M and the electrode has a linear range of 10(-4) to 3 x 10(-3) M and a response time of 180 s. Twenty L-amino acids were tested and none of them elicited responses when electrodes were exposed to 0.5 mM concentration increases over a baseline level. On the other hand, some response was observed when exposed to 18 of 20 D-amino acids varying from 2 to 200% of the corresponding D-alanine response. Electrodes were shown to have longevities of over 30 days while maintaining 85% of their original sensitivity. Electrodes showed activity over a pH of 6.2-11.7 with a maximum at 9.2 and over temperatures of 10-47 degrees C with a maximum at 37 degrees C.

Model-based optimization of a conductive matrix enzyme electrode.

A mathematical model has been developed to describe the mechanism for internal mass transfer and enzyme reaction kinetics of an amperometric conductive matrix enzyme electrode. The model is simplified and solved analytically to arrive at a representation for the response slope in the linear range as well as for the response time. This is the first time that the response time of an enzyme electrode is described by a mathematical model. Simulations give information on how the design parameters influence the performance of the electrode for a glucose oxidase catalyzed sensing reaction process. Based on this information, several designs were constructed and tested showing suitable agreement with theoretical predictions. Finally, an optimized electrode was designed and validated.

Comparing two analytical methods: minimal standards in forensic toxicology derived from information theory.

Frequently, new instrumentation and techniques are being applied to casework without an adequate knowledge of how this new technology compares to acceptable analytical procedures. Information theory provides a mathematical method to estimate the identification power of various analytical procedures and predicts if the new method produces a better or poorer confidence in the analysis than an accepted method. This paper makes comparisons based on relative informational power and discusses methods to make those estimates. With the procedures outlined in this paper, an analyst can estimate if a novel technique has the opportunity of reaching the accepted analytical methodology and, if necessary, identify places where the greatest improvement can be made to reach this equilivalence. Even though relative information theory can dismiss some proposed techniques before any analysis starts, it cannot verify if a given technique is adequate for a particular task. For that purpose, a complete procedural verification must be undertaken.