Microfluidics is a vibrant and expanding field that has the potential for solving many analytical challenges. Microfluidics shows promise to provide rapid, inexpensive, efficient, and portable diagnostic solutions that can be used in resource-limited settings. Microfluidic devices have gained immense interest as diagnostic tools for various diseases through biomarker analysis. My dissertation work focuses on developing electrokinetically operated integrated microfluidic devices for the analysis of biomarkers indicative of preterm birth risk. Preterm birth (PTB), a birth prior to 37 weeks of gestation, is the most common complication of pregnancy and the leading cause of neonatal deaths and newborn illnesses. In this dissertation, I have designed, fabricated and developed several microfluidic devices that integrate various sample preparation processes like immunoaffinity extraction, preconcentration, fluorescent labeling, and electrophoretic separation of biomarkers indicative of PTB risk. I developed microchip electrophoresis devices for separation of selected PTB biomarkers. I further optimized multiple reversed-phase porous polymer monoliths UV-polymerized in microfluidic device channels for selective retention and elution of fluorescent dyes and PTB biomarkers to facilitate on-chip labeling. Successful on-chip fluorescent labeling of multiple PTB biomarkers was reported using these microfluidic devices. These devices were further developed using a pH-mediated approach for solid-phase extraction, resulting in a ~50 fold enrichment of a PTB biomarker. Additionally, this approach was integrated with microchip electrophoresis to develop a combined enrichment and separation device that yielded 15-fold preconcentration for a PTB peptide. I also developed an immunoaffinity extraction device for analyzing PTB biomarkers directly from a human serum matrix. A glycidyl methacrylate monolith was characterized within microfluidic channels for immobilization of antibodies to PTB biomarkers. Antibody immobilization and captured analyte elution protocols were optimized for these monoliths, and two PTB biomarker proteins were successfully extracted using these devices. This approach was also integrated with microchip electrophoresis for combined extraction and separation of two PTB biomarkers in spiked human serum in <30 min. In the future, these optimized microfluidic components can be integrated into a single platform for automated immunoaffinity extraction, preconcentration, fluorescent labeling, and separation of PTB biomarkers. This integrated microfluidic platform could significantly improve human health by providing early diagnosis of PTBs.

<p>Point of care (POC) diagnosis is essential in personalized treatment to obtain effective clinical outcomes when the patient is on site. And Mass spectrometry (MS) system promotes the applications of rapid sampling ionization, which could be a tool for fast disease determination. In this thesis, a miniature MS system was firstly developed for POC tissue analysis. Lipid profile in rat organs were demonstrated. By coupling with online Paternò–Büchi (PB) reaction, fast determination of lipid C=C bond location isomers was realized. The system was applied to quantitatively analyze the change of lipid C=C location isomers between mouse normal and cancerous samples. The intensity ratio of fatty acid 18:1 (D9) and 18:1 (D11) in wild type breast tissue was calculated to be 2.881, while the ratio in tumor breast tissue was 0.667. The direct sampling-based miniature MS system is potential for POC analysis of lipid profiles and lipid biomarkers discovery.</p> <p>Secondly, an integration of paper-capillary spray and MS make it possible to analyze dried blood samples instantly in clinical laboratory and hospital. Quantitation of ratio between deuterate Phenylalanine and deuterate Tyrosine was achieved by using paper spray and paper-capillary spray MS directly, without any pretreatment of blood samples. Furthermore, these methods could generate calibration curves which enable the calculation of Phenylalanine concentration in whole human blood within 60 seconds. This disposable design is a promising application for point-of-care (POC) PKU analysis in newborn screening. </p> <p>At last, an increased in free fatty acids (FFAs) of cereals was observed during storage, and a simple and direct rice quality assessment was performed using nanoESI (Nano-Electrospray Ionization) mass spectrometry method. Six fatty acids including palmitic, oleic, and linoleic acids were compared between different rice species, growth regions and harvest years. 2D and 3D linear discriminant analysis (LDA) methods were deployed and a good sample separation was achieved. This direct sampling method of extracting FAs from rice surface combined with MS is suitable for industrial use in rapid identification for large number of samples.</p>

Palynology, the study of organic microfossils, is an important tool for improving our understanding of past environments and landscapes. Palynology provides a wealth of information from which climatic and environmental conditions can be inferred. However, inferred climatic and environmental conditions are often open to interpretation. Assumptions made about past climate conditions from pollen assemblages often rely on qualitative understanding of modern-day vegetation distributions, rather than empirical relationships. Historic anthropogenic impact on the environment must also be inferred, and assessments made as to whether vegetation changes are a result of climate change or human impact. This study seeks to address some of the questions that arise through the interpretation of pollen assemblages, by establishing empirical relationships between the geochemistry of modern pollen and climate or environmental controls. It focuses on the pollen of the climatically sensitive montane conifer Cedrus atlantica, which is distributed across the mountains of Morocco and Algeria. The study investigates aspects of modern pollen geochemistry and morphology and finds a strong relationship between the stable carbon isotope composition of modern pollen and mean annual precipitation (r2 = 0.54, p &lt;0.001) and summer precipitation (r2 = 0.63, p &lt;0.0001). Furthermore, a stronger relationship exists with aridity measured using the self-calibrating Palmer Drought Severity Index (r2 = 0.86, p &lt;0.0001), suggesting that the stable carbon isotope composition of Cedrus atlantica pollen is influenced by environmental moisture availability. The study also finds there is an increased abundance of ultraviolet absorbing compounds (UACs) in modern Cedrus atlantica pollen with increasing summer UV-B flux. This relationship was evident with samples growing in their native range (r2 = 0.84, p &lt;0.0001), but not with samples from outside this range (r2 = 0.00, p = 0.99), suggesting a possible genetic influence. Lastly, the study finds that grain size of Cedrus atlantica pollen is highly variable within and between samples, and we rule out climatic control on pollen grain size. These results suggest that quantitative relationships can be established between the geochemistry of Cedrus atlantica pollen and environmental and climatic influences. Stable carbon isotope analysis of fossil pollen could be used as a proxy for reconstruction of summer moisture availability, while analysis of UACs in fossil pollen could be used as a proxy for the reconstruction of summer UV-B flux. These proxies will enhance our understanding of climatic and environmental change in Northwest Africa and will complement existing palynological techniques for environmental and climate reconstruction. Palynology, the study of organic microfossils, is an important tool for improving our understanding of past environments and landscapes. Palynology provides a wealth of information from which climatic and environmental conditions can be inferred. However, inferred climatic and environmental conditions are often open to interpretation. Assumptions made about past climate conditions from pollen assemblages often rely on qualitative understanding of modern-day vegetation distributions, rather than empirical relationships. Historic anthropogenic impact on the environment must also be inferred, and assessments made as to whether vegetation changes are a result of climate change or human impact. This study seeks to address some of the questions that arise through the interpretation of pollen assemblages, by establishing empirical relationships between the geochemistry of modern pollen and climate or environmental controls. It focuses on the pollen of the climatically sensitive montane conifer Cedrus atlantica, which is distributed across the mountains of Morocco and Algeria. The study investigates aspects of modern pollen geochemistry and morphology and finds a strong relationship between the stable carbon isotope composition of modern pollen and mean annual precipitation (r2 = 0.54, p &lt;0.001) and summer precipitation (r2 = 0.63, p &lt;0.0001). Furthermore, a stronger relationship exists with aridity measured using the self-calibrating Palmer Drought Severity Index (r2 = 0.86, p &lt;0.0001), suggesting that the stable carbon isotope composition of Cedrus atlantica pollen is influenced by environmental moisture availability. The study also finds there is an increased abundance of ultraviolet absorbing compounds (UACs) in modern Cedrus atlantica pollen with increasing summer UV-B flux. This relationship was evident with samples growing in their native range (r2 = 0.84, p &lt;0.0001), but not with samples from outside this range (r2 = 0.00, p = 0.99), suggesting a possible genetic influence. Lastly, the study finds that grain size of Cedrus atlantica pollen is highly variable within and between samples, and we rule out climatic control on pollen grain size. These results suggest that quantitative relationships can be established between the geochemistry of Cedrus atlantica pollen and environmental and climatic influences. Stable carbon isotope analysis of fossil pollen could be used as a proxy for reconstruction of summer moisture availability, while analysis of UACs in fossil pollen could be used as a proxy for the reconstruction of summer UV-B flux. These proxies will enhance our understanding of climatic and environmental change in Northwest Africa and will complement existing palynological techniques for environmental and climate reconstruction.