Maria F. Mora

Interaction of D-Amino Acid Oxidase with Carbon Nanotubes: Implications in the Design of Biosensors

We have investigated the interaction of d-amino acid oxidase (DAAO) with single-walled carbon nanotubes (CNT) by spectroscopic ellipsometry. Dynamic adsorption experiments were performed at different experimental conditions. In addition, the activity of the enzyme adsorbed at different conditions was studied. Our results indicate that DAAO can be adsorbed to CNT at different pH values and concentrations by a combination of hydrophobic and electrostatic interactions. Considering that the highest enzymatic activity was obtained by adsorbing the protein at pH 5.7 and 0.1 mg x mL(-1), our results indicate that DAAO can adopt multiple orientations on the surface, which are ultimately responsible for significant differences in catalytic activity.

Toward Total Automation of Microfluidics for Extraterrestial In Situ Analysis

Despite multiple orbiter and landed missions to extraterrestrial bodies in the solar system, including Mars and Titan, we still know relatively little about the detailed chemical composition and quantity of organics and biomolecules in those bodies. For chemical analysis on astrobiologically relevant targets such as Mars, Europa, Titan, and Enceladus, instrumentation should be extremely sensitive and capable of analyzing a broad range of organic molecules. Microchip capillary electrophoresis (μCE) with laser-induced fluorescence (LIF) detection provides this required sensitivity and targets a wide range of relevant markers but, to date, has lacked the necessary degree of automation for spaceflight applications. Here we describe a fully integrated microfluidic device capable of performing automated end-to-end analyses of amino acids by μCE with LIF detection. The device integrates an array of pneumatically actuated valves and pumps for autonomous fluidic routing with an electrophoretic channel. Operation of the device, including manipulation of liquids for sample pretreatment and electrophoretic analysis, was performed exclusively via computer control. The device was validated by mixing of laboratory standards and labeling of amino acids with Pacific Blue succinimidyl ester followed by electrophoretic analysis. To our knowledge, this is the first demonstration of completely automated end-to-end μCE analyses on a single, fully integrated microfluidic device.

Lab-on-a-robot: Integrated microchip CE, power supply, electrochemical detector, wireless unit, and mobile platform

In this paper, the fabrication of a wireless mobile unit containing an electrochemical detection module and a 3‐channel high‐voltage power supply (HVPS) designed for microchip CE is described. The presented device consists of wireless global positioning system controlled robotics, an electrochemical detector utilizing signal conditioning analog circuitry and a digital feedback range controller, a HVPS, an air pump, and a CE microchip. A graphical user interface (LabVIEW) was also designed to communicate wirelessly with the device, from a distant personal computer communication port. The entire device is integrated and controlled by digital hardware implemented on a field programmable gate array development board. This lab‐on‐a‐robot is able to navigate to a global position location, acquire an air sample, perform the analysis (injection, separation, and detection), and send the data (electropherogram) to a remote station without exposing the analyst to the testing environment.

Microchip capillary electrophoresis instrumentation for in situ analysis in the search for extraterrestrial life.

The search for signs of life on extraterrestrial planetary bodies is among NASAs top priorities in Solar System exploration. The associated pursuit of organics and biomolecules as evidence of past or present life demands in situ investigations of planetary bodies for which sample return missions are neither practical nor affordable. These in situ studies require instrumentation capable of sensitive chemical analyses of complex mixtures including a broad range of organic molecules. Instrumentation must also be capable of autonomous operation aboard a robotically controlled vehicle that collects data and transmits it back to Earth. Microchip capillary electrophoresis (μCE) coupled to laser‐induced fluorescence (LIF) detection provides this required sensitivity and targets a wide range of relevant organics while offering low mass, volume, and power requirements. Thus, this technology would be ideally suited for in situ studies of astrobiology targets, such as Mars, Europa, Enceladus, and Titan. In this review, we introduce the characteristics of these planetary bodies that make them compelling destinations for extraterrestrial astrobiological studies, and the principal groups of organics of interest associated with each. And although the technology we describe here was first developed specifically for proposed studies of Mars, by summarizing its evolution over the past decade, we demonstrate how μCE‐LIF instrumentation has become an ideal candidate for missions of exploration to all of these nearby worlds in our Solar System.

Getting started with open-hardware: Development and control of microfluidic devices

Understanding basic concepts of electronics and computer programming allows researchers to get the most out of the equipment found in their laboratories. Although a number of platforms have been specifically designed for the general public and are supported by a vast array of on‐line tutorials, this subject is not normally included in university chemistry curricula. Aiming to provide the basic concepts of hardware and software, this article is focused on the design and use of a simple module to control a series of PDMS‐based valves. The module is based on a low‐cost microprocessor (Teensy) and open‐source software (Arduino). The microvalves were fabricated using thin sheets of PDMS and patterned using CO2 laser engraving, providing a simple and efficient way to fabricate devices without the traditional photolithographic process or facilities. Synchronization of valve control enabled the development of two simple devices to perform injection (1.6 ± 0.4 μL/stroke) and mixing of different solutions. Furthermore, a practical demonstration of the utility of this system for microscale chemical sample handling and analysis was achieved performing an on‐chip acid–base titration, followed by conductivity detection with an open‐source low‐cost detection system. Overall, the system provided a very reproducible (98%) platform to perform fluid delivery at the microfluidic scale.

Low-Temperature Microchip Nonaqueous Capillary Electrophoresis of Aliphatic Primary Amines: Applications to Titan Chemistry

We demonstrate microchip nonaqueous capillary electrophoresis (μNACE) analysis of primary aliphatic amines (C1-C18) in ethanol down to -20 °C as a first step in adapting microfluidic protocols for in situ analysis on Titan. To our knowledge, this is the first report of a nonaqueous separation at -20 °C on-chip. Limits of detection (LODs) ranged from 1.0 nM to 2.6 nM, and we identified several primary amines ranging in length from C2 to C16 in Titan aerosol analogue (tholin) samples; new amines were also detected in a tholin sample exposed to oxygen and liquid water. This preliminary work validates the sensitivity and efficacy of microfluidic chemical analysis of complex organics with relevance to Titan aerosols and surface deposits.

Implementation of microchip electrophoresis instrumentation for future spaceflight missions.

We present a comprehensive discussion of the role that microchip electrophoresis (ME) instrumentation could play in future NASA missions of exploration, as well as the current barriers that must be overcome to make this type of chemical investigation possible. We describe how ME would be able to fill fundamental gaps in our knowledge of the potential for past, present, or future life beyond Earth. Despite the great promise of ME for ultrasensitive portable chemical analysis, to date, it has never been used on a robotic mission of exploration to another world. We provide a current snapshot of the technology readiness level (TRL) of ME instrumentation, where the TRL is the NASA systems engineering metric used to evaluate the maturity of technology, and its fitness for implementation on missions. We explain how the NASA flight implementation process would apply specifically to ME instrumentation, and outline the scientific and technology development issues that must be addressed for ME analyses to be performed successfully on another world. We also outline research demonstrations that could be accomplished by independent researchers to help advance the TRL of ME instrumentation for future exploration missions. The overall approach described here for system development could be readily applied to a wide range of other instrumentation development efforts having broad societal and commercial impact.

Enhanced Resolution of Chiral Amino Acids with Capillary Electrophoresis for Biosignature Detection in Extraterrestrial Samples

Amino acids are fundamental building blocks of terrestrial life as well as ubiquitous byproducts of abiotic reactions. In order to distinguish between amino acids formed by abiotic versus biotic processes it is possible to use chemical distributions to identify patterns unique to life. This article describes two capillary electrophoresis methods capable of resolving 17 amino acids found in high abundance in both biotic and abiotic samples (seven enantiomer pairs d/l-Ala, -Asp, -Glu, -His, -Leu, -Ser, -Val and the three achiral amino acids Gly, β-Ala, and GABA). To resolve the 13 neutral amino acids one method utilizes a background electrolyte containing γ-cyclodextrin and sodium taurocholate micelles. The acidic amino acid enantiomers were resolved with γ-cyclodextrin alone. These methods allow detection limits down to 5 nM for the neutral amino acids and 500 nM for acidic amino acids and were used to analyze samples collected from Mono Lake with minimal sample preparation.

Analysis of thiols by microchip capillary electrophoresis for in situ planetary investigations

The detection of thiols on extraterrestrial bodies could provide evidence for life, as well as a host of potential prebiological or abiological processes. Here, we report a novel protocol to analyze organic thiols by microchip CE with LIF detection. Thiols were labeled with Pacific Blue C5 maleimide and analyzed by MEKC. The separation buffer consisted of 15 mM tetraborate pH 9.2 and 25 mM SDS. The optimized method provided LODs ranging from 1.4 to 15 nM. The method was validated using samples collected from geothermal pools at Hot Creek Gorge, California, which were found to contain 2‐propanethiol and 1‐butanethiol in the nanomolar concentration range. These samples serve as chemical analogues to material potentially present in the reducing environment of primitive Earth and also at sulfurous regions of Mars. Hence, the protocol developed here enables highly sensitive thiol analysis in samples with complexity comparable to that expected in astrobiologically relevant extraterrestrial settings. This new protocol could be readily added to the existing suite of microfluidic chemical analyses developed for in situ planetary exploration; all that is required is the incorporation of two new reagents to the payload of an existing instrument concept.

Investigating Protein Adsorption via Spectroscopic Ellipsometry

In this chapter, the basic concepts behind ellipsometry and spectroscopic ellipsometry are discussed along with some instrument details. Ellipsometry is an optical technique that measures changes in the reflectance and phase difference between the parallel (R P) and perpendicular (R S) components of a polarized light beam upon reflection from a surface. Aside from providing a simple, sensitive, and nondestructive way to analyze thin films, ellipsometry allows dynamic studies of film growth (thickness and optical constants) with a time resolution that is relevant to biomedical research. The present chapter intends to introduce ellipsometry as an emerging but highly promising technique, that is useful to elucidate the interactions of proteins with solid surfaces. In this regard, particular emphasis is placed on experimental details related to the development of biomedically relevant conjugated surfaces. Results from our group related to adsorption of proteins to nanostructured materials, as well as results published by other research groups, are discussed to illustrate the advantages and limitations of the technique.