Lorraine M. Siperko

SERS as a bioassay platform: fundamentals, design, and applications

Bioanalytical science is experiencing a period of unprecedented growth. Drivers behind this growth include the need to detect markers central to human and veterinary diagnostics at ever-lower levels and greater speeds. A set of parallel arguments applies to pathogens with respect to bioterrorism prevention and food and water safety. This tutorial review outlines our recent explorations on the use of surface enhanced Raman scattering (SERS) for detection of proteins, viruses, and microorganisms in heterogeneous immunoassays. It will detail the design and fabrication of the assay platform, including the capture substrate and nanoparticle-based labels. The latter, which is the cornerstone of our strategy, relies on the construction of gold nanoparticles modified with both an intrinsically strong Raman scatterer and an antibody. This labelling motif, referred to as extrinsic Raman labels (ERLs), takes advantage of the well-established signal enhancement of scatterers when coated on nanometre-sized gold particles, whereas the antibody imparts antigenic specificity. We will also examine the role of plasmon coupling between the ERLs and capture substrate, and challenges related to particle stability, nonspecific adsorption, and assay speed.

Colorimetric-solid phase extraction method for trace level determination of arsenite in water

This paper introduces a method for the determination of inorganic arsenite [As(III)] in water at low μg L(-1) by a sorption-photometric method known as colorimetric-solid phase extraction (C-SPE). The method relies on the selective extraction and concentration of an analyte on a reagent-impregnated SPE membrane, followed by direct detection of the extracted colored complex by a handheld diffuse reflectance spectrophotometer (DRS) operating in the visible spectral region. The well-established chemistry of the classic redox titrimetric method for molecular iodine (I(2)) standardization by arsenious oxide (As(2)O(3)) serves as the basis for this analysis. I(2), which is added to the aqueous sample in an excess with respect to the analyte, serves as a colorimetric indicator. The arsenite-iodine reaction is rapid, allowing an exact volume of analyte solution to be immediately passed through an SPE membrane via a syringe after mixing with the indicator. An SPE membrane that is impregnated with the complexing agent poly(vinyl-pyrrolidone) (PVP) serves to complex and concentrate excess I(2) not consumed by the As(III) analyte. The amount of complexed I(2) is determined by a DRS reading directly on the membrane surface. The spectrophotometric measurement can be made in a few seconds, with a total sample workup and readout time of ∼ 1-2 min. The limit of detection (LOD) for this determination is below 10 μg L(-1). The potential effectiveness of the method for the analysis of spiked tap water and surface water is examined, and results from preliminary interference studies are described. The work herein also shows that by applying the principles of negligible depletion (ND), the analytical procedure could be simplified by eliminating the need to pass an exact volume of a sample through the impregnated membrane as long as it exceeds the predetermined minimum volume.

Design, Certification, and Deployment of the Colorimetric Water Quality Monitoring Kit (CWQMK)

In August 2009, an experimental water quality monitoring kit based on Colorimetric Solid Phase Extraction (CSPE) technology was delivered to the International Space Station (ISS) aboard STS-128/17A. The kit, called the Colorimetric Water Quality Monitoring Kit (CWQMK), was developed by a team of scientists and engineers from NASA s Habitability and Environmental Factors Division in the Space Life Sciences Directorate at Johnson Space Center, the Wyle Integrated Science and Engineering Group in Houston, Texas, the University of Utah, and Iowa State University. The CWQMK was flown and deployed as a Station Development Test Objective (SDTO) experiment on ISS. The goal of the SDTO experiment was to evaluate the acceptability of CSPE technology for routine water quality monitoring on ISS. This paper provides an overview of the SDTO experiment, as well as a detailed description of the CWQMK hardware and a summary of the testing and analysis conducted to certify the CWQMK for use on ISS. The results obtained from the SDTO experiment are also reported and discussed in detail.

In-Flight Water Quality Monitoring on the International Space Station (ISS): Measuring Biocide Concentrations with Colorimetric Solid Phase Extraction (CSPE)

The colorimetric water quality monitoring kit (CWQMK) was delivered to the International Space Station (ISS) on STS-128/17A and was initially deployed in September 2009. The kit was flown as a station development test objective (SDTO) experiment to evaluate the acceptability of colorimetric solid phase extraction (CSPE) technology for routine water quality monitoring on the ISS. During the SDTO experiment, water samples from the U.S. water processor assembly (WPA), the U.S. potable water dispenser (PWD), and the Russian system for dispensing ground-supplied water (SVO-ZV) were collected and analyzed with the CWQMK. Samples from the U.S. segment of the ISS were analyzed for molecular iodine, which is the biocide added to water in the WPA. Samples from the SVOZV system were analyzed for ionic silver, the biocide used on the Russian segment of the ISS. In all, thirteen in-flight analysis sessions were completed as part of the SDTO experiment. This paper provides an overview of the experiment and reports the results obtained with the CWQMK. The forward plan for certifying the CWQMK as operational hardware and expanding the capabilities of the kit are also discussed.

Design, characterization, and integration of nanometric objects with chip-scale platforms for disease diagnosis

Nanomaterials (e.g., metal nanoparticles) are playing increasingly important roles in disease detection. This emergence arises from the need to detect markers and pathogens at ever-lower levels in human and veterinary diagnostics, homeland security, and food and water. This paper reviews our recent work using surface enhanced Raman scattering for detection of proteins, viruses, and microorganisms in heterogeneous immunoassays. It describes the assay platform, which consists of an antibody-modified capture substrate and gold nanoparticle-based label. The latter draws on the ability to reproducibly construct gold nanoparticles modified with a monolayer of an intrinsically strong Raman scatterer that is coated with a layer of antibodies. This construct, referred to as an extrinsic Raman label, exploits both the signal enhancement of scatterers when coated on nanometer-sized gold particles and the antigenic binding specificity of the immobilized antibody layer. Issues related to nonspecific adsorption, particle stability, and measurement reproducibility are also discussed.

Nanomaterial strategies for immunodetection

Metallic nanoparticles are playing increasingly important roles in biodiagnostic platforms. This emergence reflects the need to detect disease indicating entities at increasingly lower levels in human and veterinary diagnostics, homeland security, and food and water safety. To establish this perspective, this paper overviews our recent work using surface enhanced Raman scattering for detection of proteins, viruses, and microorganisms in heterogeneous immunoassays. It describes the assay platform, which is comprised of an antibody-modified capture substrate and gold nanoparticle-based label. The latter draws on the ability to reproducibly construct gold nanoparticles modified with a monolayer of an intrinsically strong Raman scatterer that is then coated with a layer of antibodies. This construct, referred to as an extrinsic Raman label, takes advantage of the signal enhancement of scatterers when coated on nanometer-sized gold particles and the antigenic binding specificity of the immobilized antibody layer. Challenges related to nonspecific adsorption, particle stability, and measurement reproducibility are also briefly examined.

Colorimetric Solid Phase Extraction for the Measurement of Total I (Iodine, Iodide, and Triiodide) in Spacecraft Drinking Water

An experimental drinking water monitoring kit for the measurement of iodine and silver(I) was recently delivered to the International Space Station (ISS). The kit is based on Colorimetric Solid Phase Extraction (CSPE) technology, which measures the change in diffuse reflectance of indicator disks following exposure to a water sample. To satisfy additional spacecraft water monitoring requirements, CSPE has now been extended to encompass the measurement of total I (iodine, iodide, and triiodide) through the introduction of an oxidizing agent, which converts iodide and triiodide to iodine, for measurement using the same indicator disks currently being tested on ISS. These disks detect iodine, but are insensitive to iodide and triiodide. We report here the operational considerations, design, and ground-based performance of the CSPE method for total I. The results demonstrate that CSPE technology is poised to meet NASAs total I monitoring requirements.

Colorimetric-solid phase extraction technology for water quality monitoring: Evaluation of C-SPE and debubbling methods in microgravity

Colorimetric-solid phase extraction (C-SPE) is being developed as a method for in-flight monitoring of spacecraft water quality. C-SPE is based on measuring the change in the diffuse reflectance spectrum of indicator disks following exposure to a water sample. Previous microgravity testing has shown that air bubbles suspended in water samples can cause uncertainty in the volume of liquid passed through the disks, leading to errors in the determination of water quality parameter concentrations. We report here the results of a recent series of C-9 microgravity experiments designed to evaluate manual manipulation as a means to collect bubble-free water samples of specified volumes from water sample bags containing up to 47% air. The effectiveness of manual manipulation was verified by comparing the results from C-SPE analyses of silver(I) and iodine performed in-flight using samples collected and debubbled in microgravity to those performed on-ground using bubble-free samples. The ground and flight results showed excellent agreement, demonstrating that manual manipulation is an effective means for collecting bubble-free water samples in microgravity.