Call number: LD2668 .T4 1957 C45 / Master of Science

Conducting polymers have emerged as a promising material for optoelectronics and chemical sensing application. Polyaniline (PANI) is a conductive polymer which can be easily functionalized to be specific for various biomolecules and has ideal sensor characteristics. The protonation and deprotonation of the polyaniline’s backbone by derivatization can result in color and conductive change responses. This makes it ideal for the construction of a real time, naked eye sensor. Derivatized polyaniline has previously been reported as a colorimetric sensor in solution. We plan to create a more practical sensor by synthesizing hydroxyl functionalized polyaniline thin films. In this study, we designed a process to functionalize polyaniline and deposit it as a thin film on quartz or silicon substrate via a dip coating process. To demonstrate the use of derivatized PANI in biosensing applications, derivatized and underivatized PANI thin films were treated with solutions of L-aspartic (Asp) acid at concentrations ranging from 10-8 mM to 103 mM and monitored utilizing UV-Vis spectroscopy. We found that the derivatized thin films change from deep blue to green color upon addition of Asp solution and showed a decrease in the characteristic quinoid ring peak at 600nm and the appearance of a new polaron peak at 425nm. The underivatized PANI films showed no colorimetric response indicating the hydroxyl functionalized PANI films are a more ideal material for a biosensing and naked eye detection. The polyaniline derivative was characterized using FT-IR spectroscopy, 1H NMR spectroscopy, UV-VIS spectroscopy, and Scanning Electron Microscopy. Additionally, conductivity studies were utilized to explore the material’s effectiveness as an electronic sensor using a 4-point probe to measure resistance.

Este estudo abrange as bases teóricas, com ênfase maior nos temas afins à colorimetria, dos requisitos da NBR 15936-1, de 2011, criada pela ABNT/ONS-027 Tecnologia Gráfica. A norma tem como objetivo \"...especifica[r] os requisitos para um provedor que seja capaz de produzir impressos a partir de arquivos digitais padronizados, simulando uma condição de impressão pública e aceita em todo o mundo, dentro das tolerâncias especificadas nesta Norma...e aplica-se aos processos de recepção de arquivos digitais, execução de provas digitais e processos de impressão / Thisstudycovered the theoreticalconcepts, withemphasisonthoserelatedto colorimetry, involving the requirements of NBR 15936-1, from 2011, with tile \"Graphictechnology - Quality in reproductionprocessPart 1: Requirements\", published by ABNT/ONS-027 - Graphic Technology. The standard aimsto \"specify ... the requirements for a providerthat is capable of producingstandardizedprintedfrom digital files, simulating a publicprintingcondition, accepted world wide, within the tolerancesspecified in this standard and...appliesto the processes of receiving digital files, implementation of digital proofsandprintingwithor web offset processes.\"

Este estudo abrange as bases teóricas, com ênfase maior nos temas afins à colorimetria, dos requisitos da NBR 15936-1, de 2011, criada pela ABNT/ONS-027 Tecnologia Gráfica. A norma tem como objetivo \"...especifica[r] os requisitos para um provedor que seja capaz de produzir impressos a partir de arquivos digitais padronizados, simulando uma condição de impressão pública e aceita em todo o mundo, dentro das tolerâncias especificadas nesta Norma...e aplica-se aos processos de recepção de arquivos digitais, execução de provas digitais e processos de impressão / Thisstudycovered the theoreticalconcepts, withemphasisonthoserelatedto colorimetry, involving the requirements of NBR 15936-1, from 2011, with tile \"Graphictechnology - Quality in reproductionprocessPart 1: Requirements\", published by ABNT/ONS-027 - Graphic Technology. The standard aimsto \"specify ... the requirements for a providerthat is capable of producingstandardizedprintedfrom digital files, simulating a publicprintingcondition, accepted world wide, within the tolerancesspecified in this standard and...appliesto the processes of receiving digital files, implementation of digital proofsandprintingwithor web offset processes.\"

<p>Current techniques for nucleic acid analysis often involve extensive sample preparation that requires skilled personnel and multiple purification steps. In this dissertation, we introduce an on-chip, isothermal, post-hybridization labeling and signal amplification technique that can directly interrogate unmodified DNA and RNA samples on a microarray format, eliminating the need for microarray sample pre-processing. </p><p>We name this technique Surface Initiated Enzymatic Polymerization (SIEP), where we exploit the ability of a template independent DNA polymerase called Terminal Deoxynucleotidyl Transferase (TdT) to catalyze the formation of long single-stranded DNA (ssDNA) chain from the 3'-end of a short DNA primer, which is tethered on the surface, and TdT's ability to incorporate unnatural reporter nucleotides, such as fluorescent nucleotides. We hypothesize that polymerization of a long ssDNA chain while incorporating multiple fluorescent nucleotides on target DNA or RNA hybridized to probe printed on a surface will provide a simple and powerful, isothermal method for on-chip labeling and signal amplification. </p><p>We developed the SIEP methodology by first characterizing TdT biochemical reaction to polymerize long homopolymer ssDNA (> 1000 bases) starting from the 3'-OH of ten bases oligonucleotides. We found that the preferred monomers (deoxynucleotide, dNTP) are dATP and dTTP, and that the length of the ssDNA extension is determined by the ratio of input monomer (dNTP) to initiator (short oligonucleotides). We also investigated TdT's ability to incorporate fluorescent dNTPs into a ssDNA chain by examining the effect of the molar ratios of fluorescent dNTP to natural dNTP on the initiation efficiency, the degree of fluorophore incorporation, the length and the polydispersity of the polymerized DNA strand. These experiments allowed us to incorporate up to ~50 fluorescent Cy3-labeled dNTPs per kilobase into a ssDNA chain. With the goal of using SIEP as an on-chip labeling method, we also quantified TdT mediated signal amplification on the surface by immobilizing ssDNA oligonucleotide initiators on a glass surface followed by SIEP of DNA. The incorporation of multiple fluorophores into the extended DNA chain by SIEP translated to a up to ~45 fold increase in signal amplification compared to the incorporation of a single fluorophore.</p><p>SIEP was then employed to detect hybridization of DNA (25 bases), short miRNA (21 bases) and long mRNA (1400 bases) by the post-hybridization, on-chip polymerization of fluorescently labeled ssDNA that was grown from the 3'-OH of hybridized target strands. A dose-response curve for detection of DNA hybridization by SIEP was generated, with a ~1 pM limit of detection (LOD) and a 2-log linear dynamic range while the detection of short miRNA and fragmented mRNA targets resulted in ~2 pM and ~10 pM LOD, respectively with a 3-log linear dynamic range.</p><p>We further developed SIEP for colorimetric detection by exploiting the presence of negatively charged phosphate backbone on the surface as target DNA or RNA hybridizes on the immobilized probe. The net negative charge on the surface is further increased by TdT catalyzed polymerization of long ssDNA. We then used positively charged gold nanoparticles as reporters, which can be further amplified through electroless metallization, creating DNA spots that are visible by eye. We observed an increase of 100 fold in LOD due to SIEP amplification.</p><p>Overall, we demonstrated the use of SIEP methodology to label unmodified target DNA and RNA on chip, which can be detected through fluorescence signal or colorimetric signal of metallized DNA spots. This methodology is straightforward and versatile, is compatible with current microarray technology, and can be implemented using commercially available reagents.</p> / Dissertation

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Problems associated with instruments used to measure low levels of NO₂ include non-specificity, low sensitivity, and an uncharacterized dependence on environmental conditions. MDA Scientific has recently introduced a continuous colorimetric tape technique (chemcassette) to monitor for pollutant gases. Advantages of this technology include ease of use, fast-response alarm capability, and adaptability to measure different pollutants. This research characterized and compared chemcassette performance for NO₂ with other low-level NO₂ monitoring technologies including the luminox LMA-3 monitor and the EPA reference CSI chemiluminescnt monitor. The test protocol was based on EPA procedures. The chemcassette was found to be sensitive to temperature and relative humidity effects. The chemiluminescent monitor had the best overall performance. The luminox monitor displayed temperature dependence. Variation in stability of the chemcassette optical system, tape paper uniformity, and possible interference from ozone were characterized. (Abstract shortened with permission of author.)

Epidemiological studies have linked exposure to elevated levels of airborne particulate matter with increased incidences of several types of respiratory disease, hospital admissions and morbidity. Millions of tons of airborne particulate matter are generated and released into the atmosphere each year. However, particulate matter resulting from the combustion of fuel oil and coal are of particular concern, because they are generally composed of small particles that can easily penetrate deep into the lungs, and can contain significant concentrations of toxic transition metals, such as zinc, iron and vanadium. Pulmonary toxicity (i.e. damage caused to lung tissues) of particulate matter is currently evaluated via time-consuming in-vivo testing, or via in-vitro testing. Compared to in-vivo testing, in-vitro testing offers significant advantages in terms of time savings and sample throughput. Unfortunately, the number of in-vitro testing methods are currently very limited, and do not allow a thorough investigation of the mechanisms of particulate matter toxicity. In light of these issues, the goals of the study described here were three-fold: *To adapt several in-vitro toxicity assays currently used in other applications to use in measuring particulate matter toxicity on lung cell layers; *To use these adapted assays to quantify the toxicity of numerous types of oil and coal ashes with varying particle sizes and transition metal concentrations, and; *To use the same assays to quantify the toxicities of several transition metals found in coal and oil ashes to better understand their relative contributions to overall particulate matter toxicity. Three colorimetric in-vitro assays were chosen for adaptation, and proved effective in measuring adverse cellular response to particulate matter exposure. Particle size was shown to have a large effect on the overall cytotoxicity of particulate matter; fine (less than 2.5 μm aerodynamic diameter) particles proved substantially more toxic than coarse (larger than 2.5 μm aerodynamic diameter) particles. Dose-response experiments measuring the toxic effects of the transition metals zinc, vanadium and iron revealed that zinc was the most toxic; a concentration of 0.6 mM caused a 50% drop in cellular metabolism, compared to 3 mM and 4 mM for vanadium and iron respectively.

Thesis (Ph.D.)--Rochester Institute of Technology, 2008. / Typescript. Includes bibliographical references (leaves [331]-346).

Thesis (M.S.)--Rochester Institute of Technology, 2008. / Typescript. Includes bibliographical references (leaves 269-290).