A plastic total internal reflection-based photoluminescence device for enzymatic biosensors

Thakkar, Ishan G., author
Lear, Kevin L., advisor
Reardon, Kenneth, committee member
Collins, George, committee member
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Growing concerns for quality of water, food and beverages in developing and developed countries drive sizeable markets for mass-producible, low cost devices that can measure the concentration of contaminant chemicals in water, food, and beverages rapidly and accurately. Several fiber-optic enzymatic biosensors have been reported for these applications, but they exhibit very strong presence of scattered excitation light in the signal for sensing, requiring expensive thin-film filters, and their non-planar structure makes them challenging to mass-produce. Several other planar optical waveguide-based biosensors prove to be relatively costly and more fragile due to constituent materials and the techniques involved in their fabrication. So, a plastic total internal reflection (TIR)-based low cost, low scatter, field-portable device for enzymatic biosensors is fabricated and demonstrated. The design concept of the TIR-based photoluminescent enzymatic biosensor device is explained. An analysis of economical materials with appropriate optical and chemical properties is presented. PMMA and PDMS are found to be appropriate due to their high chemical resistance, low cost, high optical transmittance and low auto-fluorescence. The techniques and procedures used for device fabrication are discussed. The device incorporated a PMMA-based optical waveguide core and PDMS-based fluid cell with simple multi-mode fiber-optics using cost-effective fabrication techniques like molding and surface modification. Several techniques of robustly depositing photoluminescent dyes on PMMA core surface are discussed. A pH-sensitive fluorescent dye, fluoresceinamine, and an O2-sensitive phosphorescent dye, Ru(dpp) both are successfully deposited using Si-adhesive gel-based as well as HydroThane-based deposition methods. Two different types of pH-sensors using two different techniques of depositing fluoresceinamine are demonstrated. Also, the effect of concentration of fluoresceinamine-dye molecules on fluorescence intensity and scattered excitation light intensity is investigated. The fluorescence intensity to the scattered excitation light intensity ratio for dye deposition is found to increase with increase in concentration. However, both the absolute fluorescence intensity and absolute scatter intensity are found to decrease in different amounts with an increase in concentration. An enzymatic hydrogen peroxide (H2O2) sensor is made and demonstrated by depositing Ruthenium-based phosphorescent dye (Ru(dpp)3) and catalase-enzyme on the surface of the waveguide core. The O2-sensitive phosphorescence of Ru(dpp)3 is used as a transduction signal and the catalase-enzyme is used as a bio-component for sensing. The H2O2 sensor exhibits a phosphorescence signal to scattered excitation light ratio of 100±18 without filtering. The unfiltered device demonstrates a detection limit of (2.20±0.6) µM with the linear range from 200µM to 20mM. An enzymatic lactose sensor is designed and characterized using Si-adhesive gel based Ru(dpp)3 deposition and oxidase enzyme. The lactose sensor exhibits the linear range of up to 0.8mM, which is too small for its application in industrial process control. So, a flow cell-based sensor device with a fluid reservoir is proposed and fabricated to increase the linear range of the sensor. Also, a multi-channel pH-sensor device with four channels is designed and fabricated for simultaneous sensing of multiple analytes.
2013 Spring.
Includes bibliographical references.
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total internal reflection fluorescence (TIRF)
indicator dye
total internal reflection
evanescent field
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