Microfabrication Process Development for a Polymer-Based Lab-on-Chip Concept Applied in Attenuated Total Reflection Fourier Transform Infrared Spectroelectrochemistry.

Micro electro-mechanical systems (MEMS) combining sensing and microfluidics functionalities, as are common in Lab-on-Chip (LoC) devices, are increasingly based on polymers. Benefits of polymers include tunable material properties, the possibility of surface functionalization, compatibility with many micro and nano patterning techniques, and optical transparency. Often, additional materials, such as metals, ceramics, or silicon, are needed for functional or auxiliary purposes, e.g., as electrodes. Hybrid patterning and integration of material composites require an increasing range of fabrication approaches, which must often be newly developed or at least adapted and optimized. Here, a microfabrication process concept is developed that allows one to implement attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and electrochemistry on an LoC device. It is designed to spatially resolve chemical sensitivity and selectivity, which are instrumental for the detection of chemical distributions, e.g., during on-flow chemical and biological reaction chemistry. The processing sequence involves (i) direct-write and soft-contact UV lithography in SUEX dry resist and replication in polydimethylsiloxane (PDMS) elastomers as the fluidic structure; (ii) surface functionalization of PDMS with oxygen plasma, 3-aminopropyl-triethoxysilane (APTES), and a UV-curable glue (NOA 73) for bonding the fluidic structure to the substrate; (iii) double-sided patterning of silicon nitride-coated silicon wafers serving as the ATR-FTIR-active internal reflection element (IRE) on one side and the electrode-covered substrate for microfluidics on the back side with lift-off and sputter-based patterning of gold electrodes; and (iv) a custom-designed active vacuum positioning and alignment setup. Fluidic channels of 100 µm height and 600 µm width in 5 mm thick PDMS were fabricated on 2" and 4" demonstrators. Electrochemistry on-chip functionality was demonstrated by cyclic voltammetry (CV) of redox reactions involving iron cyanides in different oxidation states. Further, ATR-FTIR measurements of laminar co-flows of H2O and D2O demonstrated the chemical mapping capabilities of the modular fabrication concept of the LoC devices.

Chemical Imaging of Mass Transport Near the No-Slip Interface of a Microfluidic Device using Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy.

Mass transport in geometrically confined environments is fundamental to microfluidic applications. Measuring the distribution of chemical species on flow requires the use of spatially resolved analytical tools compatible with microfluidic materials and designs. Here, the implementation of an attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) imaging (macro-ATR) approach for chemical mapping of species in microfluidic devices is described. The imaging method is configurable between a large field of view, single-frame imaging, and the use of image stitching to build composite chemical maps. Macro-ATR is used to quantify transverse diffusion in the laminar streams of coflowing fluids in dedicated microfluidic test devices. It is demonstrated that the ATR evanescent wave, which primarily probes the fluid within ∼500 nm of the channel surface, provides accurate quantification of the spatial distribution of species in the entire microfluidic device cross section. This is the case when flow and channel conditions promote vertical concentration contours in the channel as verified by three-dimensional numeric simulations of mass transport. Furthermore, the validity of treating the mass transport problem in a simplified and faster approach using reduced dimensionality numeric simulations is described. Simplified one-dimensional simulations, for the specific parameters used herein, overestimate diffusion coefficients by a factor of approximately 2, whereas full three-dimensional simulations accurately agree with experimental results.

Scanning Aperture Approach for Spatially Selective ATR-FTIR Spectroscopy: Application to Microfluidics.

We present a novel spectroscopy accessory that can easily convert any Fourier transform infrared (FTIR) spectrometer into a fully automated mapping and assaying system. The accessory uses a multiridge attenuated total reflection (ATR) wafer as the sensing element coupled with a moving aperture that is used to select the regions of interest on the wafer. In this demonstration, the accessory is combined with a series of parallel micropatterned channels, which are positioned co-linear with the light-coupling ridges on the opposite side of the ATR wafer. The ATR spectroscopy microfluidic assay accessory (ASMAA) was used in continuous mapping mode to scan perpendicular to the ATR ridges, revealing complex but repeatable oscillations in the spectral intensities. To understand this behavior, the light path through the optical components was simulated with consideration of the aperture position, ridge-to-channel alignment, and excitation beam profile. With this approach, the simulation reproduced the experimental mapping results and provided evidence that the measurement position and area changed with the aperture position. To demonstrate the assay mode, we obtained spectra along the centerline of individual microchannels and determined noise baselines and limits of detection.

Probing Heterogeneity in Attenuated Total Reflection Surface-Enhanced Infrared Absorption Spectroscopy (ATR-SEIRAS) Response with Synchrotron Infrared Microspectroscopy.

The heterogeneity of metal island films electrodeposited on conductive metal oxide modified internal reflection elements is shown to provide a variable attenuated total reflection surface-enhanced infrared absorption spectroscopy (ATR-SEIRAS) response. A self-assembled monolayer of a ferrocene-terminated thiol monolayer (FcC11SH) was formed on the gold islands covering a single substrate, which was measured using both a conventional spectrometer and a custom-built horizontal microscope. Cyclic voltammetry and ATR-SEIRAS results reveal that the FcC11SH-modified substrate undergoes a reversible electron transfer and an associated re-orientation of both the ferrocene/ferrocenium headgroup and the hydrocarbon backbone. The magnitude of the absorption signal arising from the redox changes in the monolayer, as well as the IR signature arising from the ingress/egress of the perchlorate counterions, is shown to depend significantly on the size of the infrared beam spot when using a conventional Fourier transform infrared spectrometer. By performing equivalent measurements on a horizontal microscope, the primary cause of the differences in the signal level is found to be the heterogeneity in the density of gold islands on the conductive metal oxide.

Optimization of a Commercial Variable Angle Accessory for Entry Level Users of Electrochemical Attenuated Total Reflection Surface-Enhanced Infrared Absorption Spectroscopy (ATR-SEIRAS).

An evaluation of several experimental aspects that can optimize electrochemical attenuated total reflection surface-enhanced infrared absorption spectroscopy (ATR-SEIRAS) performance using a commercially available, specular reflection accessory is provided. A comparison of different silicon single-bounce internal reflection elements (IREs) is made with emphasis on different face-angled crystal (FAC) options. Selection of optimal angle of incidence for maximizing signal and minimizing noise is shown to require consideration of the optical throughput of the accessory, reflection losses at the crystal surfaces, and polarization effects. The benefits of wire-grid polarizers and antireflective (AR) coatings on the IREs is discussed. High signal-to-noise ratios can be achieved by omitting polarizers, using an AR-coated FAC with a larger face angle, and working at angles of incidence close to the maximum throughput angle of the accessory.

Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) Spectromicroscopy Using Synchrotron Radiation and Micromachined Silicon Wafers for Microfluidic Applications.

A custom-designed optical configuration compatible with the use of micromachined multigroove internal reflection elements (µ-groove IREs) for attenuated total reflectance Fourier transform infrared (ATR FT-IR) spectroscopy and imaging applications in microfluidic devices is described. The µ-groove IREs consist of several face-angled grooves etched into a single, monolithic silicon chip. The optical configuration permits individual grooves to be addressed by focusing synchrotron sourced IR light through a 150 µm pinhole aperture, restricting the beam spot size to a dimension smaller than that of the groove walls. The effective beam spot diameter at the ATR sampling plane is determined through deconvolution of the measured detector response and found to be 70 µm. The µ-groove IREs are highly compatible with standard photolithographic techniques as demonstrated by printing a 400 µm wide channel in an SU-8 film spin-coated on the IRE surface. Attenuated total reflection FT-IR mapping as a function of sample position across the channel illustrates the potential application of this approach for rapid prototyping of microfluidic devices.

Electrochemical ATR-SEIRAS Using Low-Cost, Micromachined Si Wafers.

Thin, micromachined Si wafers, designed as internal reflection elements (IREs) for attenuated total reflectance infrared spectroscopy, are adapted to serve as substrates for electrochemical ATR surface enhanced infrared absorption spectroscopy (ATR-SEIRAS). The 500 µm thick wafer IREs with groove angles of 35° are significantly more transparent at long mid-IR wavelengths as compared to conventional large Si hemisphere IREs. The appeal of greater transparency is mitigated by smaller optical throughput at larger grazing angles and steeper angles of incidence at the reflecting plane that reduce the enhancement factor. Through use of the potential dependent adsorption of 4-methoxypyridine (MOP) as a test system, the microgroove IRE is shown to provide relatively strong electrochemical ATR-SEIRAS responses when the angle of incident radiation is between 50 and 55°, corresponding to refracted angles through the crystal of ∼40°. The higher than expected enhancement is attributed to attenuation of the reflection loss of p-polarized light and multiple reflections within the wafer-based IRE. The micromachined IREs are shown to outperform a 25 mm radius hemisphere in terms of S/N at wavenumbers less than ca. 1400 cm-1 despite the weaker signal enhancement derived from the steeper angle incident on the IRE/sample interface. The high optical transparency of the new IREs allows the spectral observation of displaced water libration bands at ca. 730 cm-1 upon solvent replacement by adsorbed MOP. The results are highly encouraging for the further development of low-cost, Si wafer-based IREs for electrochemical ATR-SEIRAS applications.

Femtomole Infrared Spectroscopy at the Electrified Metal-Solution Interface.

Characterization of surface adsorbed species using infrared (IR) spectroscopy provides valuable information concerning interfacial chemical and physical processes. However, in situ infrared studies of surface areas approaching the IR diffraction limit, such as micrometer scale electrodes, require a hitherto unrealized means to obtain high signal-to-noise (S/N) spectra from femtomole quantities of adsorbed molecules. A major methodological breakthrough is described that couples the high brilliance of synchrotron-sourced infrared microscopy with attenuated total reflection surface enhanced infrared spectroscopy (ATR-SEIRAS). The method is shown to allow the spectral measurement of a monolayer of 4-methoxypyridine (MOP) adsorbed on a surface enhancing gold film electrode under fully operational electrochemistry conditions. A factor of 15 noise improvement is achieved with small apertures using synchrotron IR relative to a thermal IR source. The very low noise levels allow the measurement of high quality IR spectra of 2.5 fmol of molecules confined to a 125 µm2 beam spot.