Electrochemical Immunosensing of Cortisol in an Automated Microfluidic System Towards Point-of-Care Applications

This dissertation describes the development of a label-free, electrochemical immunosensing platform integrated into a low-cost microfluidic system for the sensitive, selective and accurate detection of cortisol, a steroid hormone co-related with many physiological disorders. Abnormal levels of cortisol is indicative of conditions such as Cushing’s syndrome, Addison’s disease, adrenal insufficiencies and more recently post-traumatic stress disorder (PTSD). Electrochemical detection of immuno-complex formation is utilized for the sensitive detection of Cortisol using Anti-Cortisol antibodies immobilized on sensing electrodes. Electrochemical detection techniques such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) have been utilized for the characterization and sensing of the label-free detection of Cortisol. The utilization of nanomaterial’s as the immobilizing matrix for Anti-cortisol antibodies that leads to improved sensor response has been explored. A hybrid nano-composite of Polyanaline-Ag/AgO film has been fabricated onto Au substrate using electrophoretic deposition for the preparation of electrochemical immunosening of cortisol. Using a conventional 3-electrode electrochemical cell, a linear sensing range of 1pM to 1µM at a sensitivity of 66µA/M and detection limit of 0.64pg/mL has been demonstrated for detection of cortisol. Alternately, a self-assembled monolayer (SAM) of dithiobis(succinimidylpropionte) (DTSP) has been fabricated for the modification of sensing electrode to immobilize with Anti-Cortisol antibodies. To increase the sensitivity at lower detection limit and to develop a point-of-care sensing platform, the DTSP-SAM has been fabricated on micromachined interdigitated microelectrodes (µIDE). Detection of cortisol is demonstrated at a sensitivity of 20.7µA/M and detection limit of 10pg/mL for a linear sensing range of 10pM to 200nM using the µIDE’s. A simple, low-cost microfluidic system is designed using low-temperature co-fired ceramics (LTCC) technology for the integration of the electrochemical cortisol immunosensor and automation of the immunoassay. For the first time, the non-specific adsorption of analyte on LTCC has been characterized for microfluidic applications. The design, fabrication technique and fluidic characterization of the immunoassay are presented. The DTSP-SAM based electrochemical immunosensor on µIDE is integrated into the LTCC microfluidic system and cortisol detection is achieved in the microfluidic system in a fully automated assay. The fully automated microfluidic immunosensor hold great promise for accurate, sensitive detection of cortisol in point-of-care applications.

Step and flash imprint lithography: Fabrication of patterned media for extremely high density magnetic data storage devices

Currently the data storage industry is facing huge challenges with respect to the conventional method of recording data known as longitudinal magnetic recording. This technology is fast approaching a fundamental physical limit, known as the superparamagnetic limit. A unique way of deferring the superparamagnetic limit incorporates the patterning of magnetic media. This method exploits the use of lithography tools to predetermine the areal density. Various nanofabrication schemes are employed to pattern the magnetic material are Focus Ion Beam (FIB), E-beam Lithography (EBL), UV-Optical Lithography (UVL), Self-assembled Media Synthesis and Nanoimprint Lithography (NIL). Although there are many challenges to manufacturing patterned media, the large potential gains offered in terms of areal density make it one of the most promising new technologies on the horizon for future hard disk drives. Thus, this dissertation contributes to the development of future alternative data storage devices and deferring the superparamagnetic limit by designing and characterizing patterned magnetic media using a novel nanoimprint replication process called "Step and Flash Imprint lithography". As opposed to hot embossing and other high temperature-low pressure processes, SFIL can be performed at low pressure and room temperature. Initial experiments carried out, consisted of process flow design for the patterned structures on sputtered Ni-Fe thin films. The main one being the defectivity analysis for the SFIL process conducted by fabricating and testing devices of varying feature sizes (50 nm to 1 μm) and inspecting them optically as well as testing them electrically. Once the SFIL process was optimized, a number of Ni-Fe coated wafers were imprinted with a template having the patterned topography. A minimum feature size of 40 nm was obtained with varying pitch (1:1, 1:1.5, 1:2, and 1:3). The Characterization steps involved extensive SEM study at each processing step as well as Atomic Force Microscopy (AFM) and Magnetic Force Microscopy (MFM) analysis.