Microfluidics is gaining in importance due to its wide ranging benefits and applicability in chemical and biological analysis. Although traditional microfluidic devices are created with glass or silicon based fabrication technologies, polymer based devices are gaining in popularity. Soft lithography and replica molding are techniques for the rapid prototyping of such devices, utilizing Polydimethylsiloxane (PDMS) as the dominant material. Other benefits include its low costs and ease of fabrication. Even though soft lithography is a well researched and developed fabrication process, new applications have been discovered in which the technology can be applied. Often, changes in the fabrication process are necessary for their application in other areas of research. This thesis will address several microfluidic applications using soft lithography. These areas of research include microfluidic thermometry, isoelectric focusing (IEF), and micromixers. In microfluidic thermometry, a novel thin film PDMS/Rhodamine B has been developed allowing whole-chip temperature measurements. In addition, compatibility problems between Rhodamine B and PDMS microfluidic devices were resolved. The thin film fabrication process, experimental results, and issues with its use are discussed. Future work and attempts at improving the thin film performance are also provided. IEF involves applications in which samples are separated according to its electrostatic charge. Two types of IEF applications are shown in which soft lithography has been shown to be beneficial to its development and performance. In isoelectric focusing with the use of thermally generated pH gradients, soft lithography allows for the rapid design, production and testing of different channel layouts. In general, due to PDMS insulation and overall low heat transfer rates, the temperatures detected are more gradual than those previously reported in literature. IEF using carrier ampholytes are also discussed, with preliminary results in which devices fabricated using soft lithography are compared to commercially available IEF cartridges. Its fabrication issues are discussed in detail. In micromixers, soft lithography fabrication issues and overall integration with flow mechanisms is discussed. In general it is difficult to perform mixing in the microscale due to the predominantly laminar flow and flow rate restrictions. Channel geometry is insignificant, as can be seen through numerical simulations.

Microfluidics is gaining in importance due to its wide ranging benefits and applicability in chemical and biological analysis. Although traditional microfluidic devices are created with glass or silicon based fabrication technologies, polymer based devices are gaining in popularity. Soft lithography and replica molding are techniques for the rapid prototyping of such devices, utilizing Polydimethylsiloxane (PDMS) as the dominant material. Other benefits include its low costs and ease of fabrication. Even though soft lithography is a well researched and developed fabrication process, new applications have been discovered in which the technology can be applied. Often, changes in the fabrication process are necessary for their application in other areas of research. This thesis will address several microfluidic applications using soft lithography. These areas of research include microfluidic thermometry, isoelectric focusing (IEF), and micromixers. In microfluidic thermometry, a novel thin film PDMS/Rhodamine B has been developed allowing whole-chip temperature measurements. In addition, compatibility problems between Rhodamine B and PDMS microfluidic devices were resolved. The thin film fabrication process, experimental results, and issues with its use are discussed. Future work and attempts at improving the thin film performance are also provided. IEF involves applications in which samples are separated according to its electrostatic charge. Two types of IEF applications are shown in which soft lithography has been shown to be beneficial to its development and performance. In isoelectric focusing with the use of thermally generated pH gradients, soft lithography allows for the rapid design, production and testing of different channel layouts. In general, due to PDMS insulation and overall low heat transfer rates, the temperatures detected are more gradual than those previously reported in literature. IEF using carrier ampholytes are also discussed, with preliminary results in which devices fabricated using soft lithography are compared to commercially available IEF cartridges. Its fabrication issues are discussed in detail. In micromixers, soft lithography fabrication issues and overall integration with flow mechanisms is discussed. In general it is difficult to perform mixing in the microscale due to the predominantly laminar flow and flow rate restrictions. Channel geometry is insignificant, as can be seen through numerical simulations.

Submitted by Maria Medeiros (maria.dilva1@ufcg.edu.br) on 2018-07-11T13:13:10Z No. of bitstreams: 1 JOÃO FERNANDES DUARTE NETO - DISSERTAÇÃO (PPGCEMat) 2015.pdf: 2295774 bytes, checksum: d73166400ea15685ca449c373a12dc20 (MD5) / Made available in DSpace on 2018-07-11T13:13:10Z (GMT). No. of bitstreams: 1 JOÃO FERNANDES DUARTE NETO - DISSERTAÇÃO (PPGCEMat) 2015.pdf: 2295774 bytes, checksum: d73166400ea15685ca449c373a12dc20 (MD5) Previous issue date: 2015-02-24 / Este trabalho investigou o processo de adsorção, equilibrio e cinética de argilas esmectíticas da cidade de Boa Vista (Paraíba), usadas como adsorventes de corantes com comportamentos químicos distintos - rodamina B (RB), catiônico, e alaranjado de metila (AL), aniônico - em meio aquoso. Essas argilas foram escolhidas devido às suas propriedades características do argilomineral esmectítico, que possui sítios quimicamente ativos, provocado pela substituição isomórfica e interação das arestas, tornando-o atrativo sua aplicação para fins adsortivos. As argilas utilizadas são conhecidas como bentonita chocolate e bentonita chocobofe. Esses materiais foram utilizados na forma natural e após tratamento com carbonato de sódio, ácido clorídrico e organofilização (sal quaternário de amônio PRAEPAGEN). Todos os adsorventes foram caracterizados por fluorescência de raios X, difração de raios X, análises térmicas (TG, DTG e DTA) e determinação de tamanho de partícula. Os ensaios de adsorção dos corantes foram examinados por sistema de bateladas, alterando tempo de contato, quantidade de adsorvente, concentração dos corantes e pH inicial. Para a análise do comportamento de adsorção foi utilizado espectrofotometria UV-VIS. O modelo de isoterma de Freundlich foi o que melhor se ajustou aos dados experimentais das argilas naturais e modificadas. Porém nos processos adsortivos envolvendo as argilas organofilizadas, na interação com RB, tanto o modelo de Langmuir como o de Freundlich apresentaram um bom ajuste aos dados experimentais. Os dados cinéticos de adsorção foram ajustados ao modelo de pseudo-segunda ordem (R2>0,9). Os testes de capacidades de adsorção permitiram concluir que a maior eficiência em remover a RB ocorre quando as argilas são tratadas com ácido clorídrico. A maior eficiência na remoção do AL ocorre quando as argilas estudadas foram as organofilizadas. De modo geral, as argilas esmectíticas estudadas se mostraram adsorventes adequados para a remoção do corante RB, sendo, no entanto, necessário tratamentos específicos para maximizar ou melhorar a capacidade de adsorção dos corantes RB e AL. / This study investigated the process of adsorption, equilibrium and kinetics, of smectite clays from Boa Vista of Paraíba used as chemical adsorbents of dyes with different ionic behavior in aqueous media. Smectite clays known as chocolate and chocobofe and the dyes Rhodamine B (RB) (cationic) and Methyl Orange (AL), (anionic) were used in this study. The adsorbents were characterized by X-ray fluorescence, X-ray diffraction, thermal analysis (TG, DTA and DTG) and determination of particle size. Clays have been used in natural form and after treatment with sodium carbonate, acid and organophilization (quaternary ammonium salt – PRAEPAGEN). The dye adsorption was examined by a batch system, changing: contact time, amount of adsorbent, dye concentration and initial pH. UVVIS spectrophotometry was used for the analysis of the adsorption behavior. The clays were characterized as smectite clays and the efficiency of the treatments was confirmed by X-ray fluorescence and X-ray diffraction. The model of Freundlich isotherm presented the best fit to the experimental data of natural clays. However, in the adsorptive processes involving clays organophilizated both the Langmuir model and the Freundlich model showed good fitting to the experimental data. The kinetic adsorption data were fitted to the model of pseudo-second order (R2> 0.9). The adsorption capacities test showed that the highest efficiency in removing RB occurs when the clays are treated with hydrochloric acid. The highest efficiency of the removal of AL occurs when the clays are organophilizated. In general, the smectite clays were suitable adsorbent for the removal of the dye RB. However, special treatment is necessary to maximize or improve the adsorption ability of the dyes AL and RB.

Este trabalho investigou a fotodegradação do corante Rodamina-B com o auxílio do catalisador semicondutor Dióxido de Titânio, utilizando como fonte de irradiação pulsos de 40 fs gerados por um sistema laser de Titânio - Safira. Os pulsos laser foram formatados espectralmente (modulação senoidal), gerando trens de pulsos cuja separação foi escolhida para ser ressonante com um modo Raman intenso do catalisador. A fotodegradação da Rodamina-B foi monitorada através de medidas de fluorescência excitada via absorção de dois fótons. Não observamos fotodegradação do corante mediante a irradiação com pulsos ultracurtos, mesmo na presença do catalisador, tanto para o pulso de menor duração alcançada quanto para os trens de pulsos com diferentes separações temporais. Contudo, observamos uma diminuição no espalhamento da luz de excitação devido às nanopartículas de Dióxido de Titânio. Este processo foi estudado com mais detalhes, tendo sido analisada a influência da formatação espectral senoidal no resultado da medida. A relação entre a irradiação com pulsos de femtossegundos e a quebra de aglomerados de nanopartículas foi intuída a partir dos resultados obtidos, sendo fortalecida por medidas de microscopia eletrônica de varredura de um filme das amostras. / This work investigated the photodegradation of Rhodamine-B assisted by Titanium Dioxide semiconductor catalyst, using as irradiation source a Ti:Sapphire laser system delivering 40 fs pulses. The laser pulses were spectrally shaped (sinusoidal modulation), generating pulse trains whose separation was chosen to be resonant with an intense catalysts Raman mode. The Rh-B photodegradation was monitored by two photon excited fluorescence measurements. We have not observed dye photodegradation by the ultrashort pulses irradiation, in the absence of the catalyst, neither with the shortest pulse obtained, nor with pulse trains. Nevertheless, we observed a decrease in the excitation light scattering due to the Titanium Dioxide nanoparticles. Such process was studied by analyzing the influence of sinusoidal spectral phase mask in the signal. The results indicate a relationship between femtosecond pulse irradiation and the breaking of nanoparticles clusters, which was also corroborated by scanning electron microscopy measurements.

Submitted by Diego da Silva Manoel null (diegosma@rc.unesp.br) on 2017-02-05T16:40:20Z No. of bitstreams: 1 Dissertação Diego.pdf: 1793320 bytes, checksum: ba0a17055ebb3486b214146535165ef9 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2017-02-06T18:25:08Z (GMT) No. of bitstreams: 1 manoel_ds_me_rcla.pdf: 1793320 bytes, checksum: ba0a17055ebb3486b214146535165ef9 (MD5) / Made available in DSpace on 2017-02-06T18:25:08Z (GMT). No. of bitstreams: 1 manoel_ds_me_rcla.pdf: 1793320 bytes, checksum: ba0a17055ebb3486b214146535165ef9 (MD5) Previous issue date: 2017-01-17 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O processo sol-gel permite a produção de materiais Híbridos Orgânicos/Inorgânicos para aplicações em óptica e fotônica. Materiais Híbridos de Sílica/orgânico unem propriedades da Sílica e dos polímeros orgânicos e permitem a incorporação de diversas moléculas orgânicas na matriz, possibilitando a obtenção de materiais fotoativos avançados. Destas moléculas destacam-se os corantes lasers, como a Rodamina-B, que possui comprimento de onda de emissão bem definido e alta eficiência quântica. Neste trabalho preparamos materiais híbridos de sílica/orgânico via processo sol-gel utilizando como precursores os alcóxidos de silício 3-glicidoxipropiltrimetoxisilano (GPTS) e tetraetilortosilicato (TEOS). Amostras ainda na fase de sol foram dopadas com Rodamina-B em concentrações variando de 0,01 a 5 mmol/L. A fluorescência das amostras foi caracterizada mostrando dependência entre as características da emissão e a concentração de dopante. Os espectros de fluorescência sofrem um deslocamento para a região do vermelho com o aumento da concentração do dopante, enquanto a intensidade de emissão apresenta maior valor para a amostra com concentração de 0,5 mmol/L. Também foram determinados os coeficientes de absorção óptica, que apresentaram comportamento linear com a concentração. A partir das amostras (sol) foram produzidos xerogéis com concentrações de Rodamina-B de 0,0386 a 19,3 mmol/L. Os espectros de fluorescência destes xerogéis também apresentaram deslocamento para a região do vermelho para maiores concentrações de dopante, a intensidade de emissão é máxima para o xerogel com concentração de 6,56 mmol/L. Os coeficientes de absorção óptica dos xerogéis apresentaram um comportamento linear com a concentração de dopante. O xerogel com concentração de 19,3 mmol/L de Rodamina-B apresentou emissão de laser randômico quando excitado com laser pulsado (532 nm, 8ns), fenômeno caracterizado por um estreitamento da banda de emissão devido a dímeros da Rodamina-B em 630 nm. Microscopia eletrônica de transmissão revelou a formação de partículas de Sílica com diâmetro médio de 6,3 nm distribuídas na matriz sólida. Estes resultados são de grande interesse para aplicações em fotônica. / Organic-Inorganic Hybrid Materials prepared by sol-gel process have applications in different areas such as optics and photonics. Silica/organic hybrid materials combine the properties from the silica and the organic polymers and can be doped with organic molecules with different optical properties, allowing preparation of advanced photoactive materials. In the wide range of photoactive organic materials one of interest is the Rhodamine-B, a laser dye which presents tunable emission and high quantum yield. In this work we have produced hybrid silica/organic samples derived from 3-glycidoxypropyltrimethoxysilane (GPTS) and tetraethylorthosilicate (TEOS) by sol-gel process. Silica/organic sols were doped with Rhodamine-B concentrations ranging from 0.01 to 5 mmol/L. The fluorescence spectra of the samples were characterized and a relationship between emission characteristics and dopant concentration has been observed. A red shift in the wavelength emission was observed, as the dopant concentration increased and the maximum emission intensity was achieved for the sample with concentration of 0.5mmol/L. Were determined the optical absorption coefficients that showed a linear relation with the concentration. The sol samples were dried and xerogels were obtained and the new concentrations of Rhodamine-B in the solid matrix were determined, ranging from 0.0386 to 19.3 mmol/L. The fluorescence spectra of the xerogels were characterized, showing a red shift as the dopant concentration increased and the maximum emission intensity was achieved for the xerogel with concentration of 6.56 mmol/L. The optical absorption coefficients of these xerogels showed a linear behavior with the dopant concentration. Random laser action for the xerogel with concentration of 19.3 mmol/L was measured using an pulsed laser (532 nm, 8ns), and characterized by a narrowing emission of the Rhodamine-B dimmers emission at 630 nm. Transmission electron microscopy revealed silica particles with 6.3 nm average diameter in the solid matrix. These results are very important for applications in photonics.

A novel electrochemical genosensor with modified graphite with poly(4-aminophenol) has been constructed for detection of neuroblastoma, a malignant tumor originating from embryonic precursor cells of the sympathetic nervous system and associated with the amplification MYCN oncogene. The produced genosensor exhibited distinct electric and morphological properties using rhodamine b, specie able to bind with DNA duplex, as indicator of the hybridization process. The detection limit was evaluated to be 0.47 umol.L-1 (N=3) and showed very high selectivity for the complementary DNA using serum sample. This DNA sensing platform was successfully applied to detect the MYCN, an important biomarker for neuroblastoma / Um novo genossensor eletroquímico de grafite modificado com poli (4-aminofenol) foi construído para a detecção de neuroblastoma, um tumor maligno originário a partir de células precursoras embrionárias do sistema nervoso simpático, e associado com a amplificação do oncogene MYCN. O genossensor produzido exibiu propriedades elétricas e morfológicas distintas, utilizando rodamina B, espécie capaz de se ligar com a fita dupla de DNA, como indicador do processo de hibridação. O limite de detecção obtido foi de 0,47 μmol.L-1 (N = 3) e mostrou maior seletividade para o DNA complementar, utilizando amostras de soro. Esta plataforma de detecção de DNA foi aplicada com sucesso para detectar a MYCN, um biomarcador importante para o neuroblastoma / Mestre em Genética e Bioquímica

A series of picosecond excite-probe experiments was performed on various concentrations of aqueous and ethanolic solutions of rhodamine B in order to determine the existence of dimerization in those solutions. The goals of the research presented in this dissertation were twofold. Initially, various techniques of time-resolved spectroscopy were to be employed to further characterize the ground and excited-state molecular properties of the aqueous RB dimer. The information obtained, and the techniques developed in that study would then be utilized in an effort to secure evidence which would support or refute the claims of rhodamine B dimerization in an ethanolic solution.

Gene regulation by transcription factors are one of the key processes that are important to sustain all kinds of life. In the prokaryote Escherichia coli this has shown to especially crucial. The operator sequence to which these transcription factors bind to are very small in comparison to the whole genome of E. coli, thus the question becomes how these proteins can find these sequences quickly. One particularly well-studied transcription factor in this regard is the lac repressor. It has been shown that this transcription factors finds its operators faster than the limit of three dimensional diffusion. The leading model for how the repressor does that is facilitated diffusion and this model has gained more experimental evidence, particularly using single-molecule fluorescence microscopy. This study aimed at measuring the unspecific binding time between the lac repressor and DNA in vivo, but in the end the project evolved to trying to establish a single-molecule tracking assay of the repressor in vivo. In this study a mutant of the repressor was expressed and purified, labelled with a synthetic fluorophore, electroporated into E. coli and tracking was performed under a microscope. One of the three types of experiments were partially analysed with an image analysis software. Unfortunately, analysis was not completed for all experiments which made it difficult to compare the results. In the end the data was compared by eye while also using the results from image analysis. With slight optimism it can be concluded that the assay worked, but it needs more development.

To effectively design systems of microchannels it is necessary for scientists and engineers to understand thermal transport characteristics of microchannels. To experimentally determine the convective heat transfer coefficient of microchannels it is necessary to measure both the bulk and surface temperature fields. This investigation aims to develop a technique, named Total Internal Reflection Fluorescent Micro-Thermometry (TIR-FMT), to measure the temperature of water within several hundred nanometers of a wall--effectively, the surface temperature of the wall. In TIR-FMT, an evanescent-wave is generated in the water near the wall. The intensity of this evanescent-wave decays exponentially with distance from the wall. A fluorophore if illuminated by the evanescent-wave can absorb a photon. Excited fluorophores subsequently emit red-shifted photons, which are called fluorescence. The probability of a fluorescent emission is temperature-dependent. Therefore, by monitoring the intensity of the fluorescence a correlation can be made to the temperature of the region of illumination. Using the TIR-FMT technique the temperature dependence of the fluorescence intensity from buffered fluorescein (pH=9.2) was determined to be 1.35%/C. TIR-FMT can be used to measure the temperature of a fluorophore solution within 600 nm of a wall across a temperature range of 12.5-55C. The average uncertainties (95% confidence) of the temperature measured was determined to be 2.3C and 1.5C for a single 1.5x1.5 and #956;m pixel and the entire 715x950 and #956;m viewfield. By spatial averaging, average uncertainties of 2.0C and 1.8C were attained with spatial resolutions of 16x16 and 100x100 and #956;m, respectively.

M. Tech. (Department of Chemistry, Faculty of Applied and Computer Sciences), Vaal University of Technology. / The ligand 2, 2-dithiobisbenzothiazole and it metal complexes have been a subject of interest in various fields but they have found to exhibit remarkable and prevalent biological and pharmacological activities. The ligand tends to coordinate to complexes through the sulfur atom and hence the metal-sulphide bond are good precursor to generate metal sulfide nanoparticles using single-source precursor route. The complexes are generally prepared by reflux for 1 to 2 hours depending on the solvent used to produce very stable solid products and some form in crystalline form. All the prepared nickel and cadmium complexes were characterized using techniques such as elemental analyzer, IR, 13C NMR spectroscopy and thermogravimatric analysis. The data obtained from the spectroscopic analysis was consistent of the coordination of the ligand with the metal ions through the sulphur atoms of the 2,2-dithiobisbenzothiazole moiety. The thermal analysis of the prepared complexes gave a final residue of metal sulphide for both metal complexes. Characterization techniques showed the formation of bidentate complexes for both nickel complex and cadmium complex. The prepared complexes were then used to synthesize metal sulphide nanoparticles .The nanoparticles were prepared by thermal decomposition method of the single source precursor in a solution of oleylamine (OLA). Two different parameters were investigated temperature and time to study their effect on the size and shape of the nanoparticles. The synthesized nanoparticles were characterized using techniques such as UV-Vis spectroscopy, photoluminescence spectroscopy, and X-ray diffraction analysis and transmission electron microscopy. The temperatures of the reaction have a significant effect on the rate of the reaction that will affect the size and shape of the nanoparticles. This effect was confirmed by the optical properties of the synthesized nanoparticles prepared at different reaction temperatures. The spectra shows that absorption maximum and band edge shift to lower wavelength as the temperature of reaction was progressively increased. This trend is associated to the decrease in particles size of the prepared nanoparticles. TEM images further confirmed that the particles size of the prepared nanoparticles was progressively decreased as the temperature was increased. The time of the reaction is one of the most significant factors in the synthesis of the nanoparticles. The investigation of the time of the reaction yield results that depicted that with increase in time of the reaction, the band edge increases, but relatively at short wavelength to the bulk. Hence, the band edges of the nanoparticles were blue shifted significantly to the bulk. The results show that with an increase in the time of the reaction, the nanoparticles increases in their size due to Ostwald ripening. The optimum complexes and optimum nanoparticles were used to further study their electrochemical properties using cyclic voltammetry and electrochemical impedance spectroscopy (EIS) graphs were fitted using the randles circuit and they confirm that the NiS nanoparticles GCE greatly increase the electron transfer rate, probably due to the nanostructured surface property of the NiS nanoparticles. Differential pulse voltammetry (DPV) was used to study the electrochemical behavior and the DPV showed that the current response of Rhb was higher for the optimum temperature NiS nanoparticles compared to all the materials used. There was an increase in the Rhb current response with an increase in pH and pH 7 was used as the optimum pH when Ni- complex was used as a modifier and pH 8 was used as optimum when NiS nanoparticles were used as a modifier. Effect of concentration showed that the NiS nanoparticles for the optimum temperature had a wide linear range and a low detection limit. The method has good accuracy, acceptable precision, and reproducibility. This method provides a novel electrochemical method for determination of RhB.