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Design, Development and Testing of a Fluorescence-based Microfluidics System for Uric Acid Analysis of Clinical Samples Open Access

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The work described in this dissertation involved the design, development and testing of a fluorescence-based microfluidics analytical system. Initially, a new and thorough, yet tractable, computational methodology had to be developed for estimating the performance of optical microfluidics systems. Equations are provided to trace quantitatively the flow of photons through optical components, samples and spectral-selective components, to the detectors. The new computational methodology is useful for calculating calibration curves rapidly for a very wide variety of designs. This methodology also provides the basis for both detailed understanding and improved design of microfluidic optical analytical systems. 2000 computations were performed with the new tool, and an effective system design resulted.A novel compact fluorescence analysis system for quantification of uric acid (UA) in clinical samples was designed, fabricated and tested. To perform an analysis, diluted saliva, urine or blood samples are simply placed in the new disposable functionalized thin-film holder. Two enzymes and Amplex Red were immobilized within the sample holder. A compact prototype instrument was developed. It contained an LED, a narrow-band filter and an amplified photodiode. The analysis time is 30 seconds, and the dynamic range of the system is 4 to 400 µM of UA. The calibration curve for saliva and urine was made using solutions of UA. The calibration curve for blood was obtained with spiked samples of blood. Three different types of clinical samples were collected from three subjects. Analysis with our instrument yielded UA concentrations within the expected concentration ranges. Overall, 600 UA measurements were made.

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