Frederik H. Kriel

Pillar cuvettes: capillary-filled, microliter quartz cuvettes with microscale path lengths for optical spectroscopy.

The goal of most analytical techniques is to reduce the lower limit of detection; however, it is sometimes necessary to do the opposite. High sample concentrations or samples with high molar absorptivity (e.g., dyes and metal complexes) often require multiple dilution steps or laborious sample preparation prior to spectroscopic analysis. Here, we demonstrate dilution-free, one-step UV-vis spectroscopic analysis of high concentrations of platinum(IV) hexachloride in a micropillar array, that is, pillar cuvette. The cuvette is spontaneously filled by wicking of the liquid sample into the micropillar array. The pillar height (thus, the film thickness) defines the optical path length, which was reduced to between 10 and 20 μm in this study (3 orders of magnitude smaller than in a typical cuvette). Only one small droplet (∼2 μL) of sample is required, and the dispensed volume need not be precise or even known to the analyst for accurate spectroscopy measurements. For opaque pillars, we show that absorbance is linearly related to platinum concentration (the Beer-Lambert Law). For fully transparent or semitransparent pillars, the measured absorbance was successfully corrected for the fractional surface coverage of the pillars and the transmittance of the pillars and reference. Thus, both opaque and transparent pillars can be applied to absorbance spectroscopy of high absorptivity, microliter samples. It is also shown here that the pillar array has a useful secondary function as an integrated (in-cuvette) filter for particulates. For pillar cuvette measurements of platinum solutions spiked with 6 μm diameter polystyrene spheres, filtered and unfiltered samples gave identical spectra.

Influence of Sample Volume and Solvent Evaporation on Absorbance Spectroscopy in a Microfluidic "Pillar-Cuvette".

Spectroscopic analysis of solutions containing samples at high concentrations or molar absorptivity can present practical challenges. In absorbance spectroscopy, short optical path lengths or multiple dilution is required to bring the measured absorbance into the range of the Beers Law calibration. We have previously reported an open pillar-cuvette with a micropillar array that is spontaneously filled with a precise (nL or μL) volume to create the well-defined optical path of, for example, 10 to 20 μm. Evaporation should not be ignored for open cuvettes and, herein, the volume of loaded sample and the rate of evaporation from the cuvette are studied. It was found that the volume of loaded sample (between 1 and 10 μL) had no effect on the Beers Law calibration for methyl orange solutions (molar absorptivity of (2.42 ± 0.02)× 10(4) L mol(-1) cm(-1)) for cuvettes with a 14.2 ± 0.2 μm path length. Evaporation rates of water from methyl orange solutions were between 2 and 5 nL s(-1) (30 - 40% relative humidity; 23°C), depending on the sample concentration and ambient conditions. Evaporation could be reduced by placing a cover slip several millimeters above the cuvette. Importantly, the results show that a drop-and-measure method (measurement within ∼3 s of cuvette loading) eliminates the need for extrapolation of the absorbance-time data for accurate analysis of samples.

The use of microfluidics in cytotoxicity and nanotoxicity experiments

Many unique chemical compounds and nanomaterials are being developed, and each one requires a considerable range of in vitro and/or in vivo toxicity screening in order to evaluate their safety. The current methodology of in vitro toxicological screening on cells is based on well-plate assays that require time-consuming manual handling or expensive automation to gather enough meaningful toxicology data. Cost reduction; access to faster, more comprehensive toxicity data; and a robust platform capable of quantitative testing, will be essential in evaluating the safety of new chemicals and nanomaterials, and, at the same time, in securing the confidence of regulators and end-users. Microfluidic chips offer an alternative platform for toxicity screening that has the potential to transform both the rates and efficiency of nanomaterial testing, as reviewed here. The inherent advantages of microfluidic technologies offer high-throughput screening with small volumes of analytes, parallel analyses, and low-cost fabrication.

Directed Growth of Orthorhombic Crystals in a Micropillar Array

We report directed growth of orthorhombic crystals of potassium permanganate in spatial confinement of a micropillar array. The solution is introduced by spontaneous wicking to give a well-defined film (thickness 10-15 μm; volume ∼600 nL) and is connected to a reservoir (several microliters) that continuously feeds the evaporating film. When the film is supersaturated, crystals nucleate and preferentially grow in specific directions guided by one of several possible linear paths through the pillar lattice. Crystals that do not initially conform are stopped at an obstructing pillar, branch into another permitted direction, or spontaneously rotate to align with a path and continue to grow. Microspectroscopy is able to track the concentration of solute in a small region of interest (70 × 100 μm2) near to growing crystals, revealing that the solute concentration initially increases linearly beyond the solubility limit. Crystal growth near the region of interest resulted in a sharp decrease in the local solute concentration (which rapidly returns the concentration to the solubility limit), consistent with estimated diffusion time scales (<1 s for a 50 μm length scale). The ability to simultaneously track solute concentration and control crystal orientation in nanoliter samples will provide new insight into microscale dynamics of microscale crystallization.