I. Hadamard Transform Capillary Electrophoresis for the Analysis of Biologically Active Species II. Characterization and Application of Two-Photon Activatable Proton and Radical Generators

by Braun, Kevin L.

PART I. A modified Hadamard transform has been developed and applied to the analysis of biologically active species using capillary electrophoresis. Hadamard transformations, a matrix based multiplexing technique, when coupled with a capillary electrophoresis instrument capable of rapid sample injection, provides a means to semicontinuously inject samples. The multiple injections separate, interpenetrate, and are detected as the summation of the multiple injections. Deconvolution of the multiplexed signal by multiplication with the inverse of the injection matrix yields a single injection electropherogram that exhibits improved S/N. In modified Hadamard transform capillary electrophoresis (mHTCE), an injection sequence of half the length as conventional HTCE (cHTCE) is utilized. Modifying the manner in which the raw data is manipulated before deconvolution facilitates the reduced injection sequence. When coupled with software, mHTCE can reduce the collection time for a Hadamard sequence by up to 48%. The substantial time reduction afforded by mHTCE is utilized to demonstrate the first timeresolved application of Hadamard transformations for the analysis of neurotransmitters. Additionally, mHTCE has been demonstrated as a means to improve the sensitivity for analysis of amino acids and proteins including ?-aminobutyric acid, dopamine, and enhanced green fluorescent protein (EGFP) with picomolar detection limits. Part II. Two-photon excitation provides a means to activate chemical and physical processes with high spatial resolution and improved depth penetration compared to one-photon excitation. When combined with three-dimensional lithographic microfabrication (3DLM), these advantages provide a means to fabricate complex 22 structures through radical and cationic two-photon induced polymerization (TPIP). A strategy for realizing high-fidelity microstructures is reported that considers the inherent structural limitations of acrylate monomers. Utilizing this strategy, a series of highfidelity microstructures is reported for application in microfluidic devices, microelectromechanical systems (MEMS), and microoptical devices such as photonic bandgap (PBG) crystals. Improved periodicity is reported here for f.c.c. PBG crystals compared to earlier examples through addition of micromechanical supports that provide increased strength to the high-aspect ratio crystals. To extend TPIP to cationic polymerization, a series of two-photon activatable photoacid generators has been developed. The new PAGs exhibit one to two orders of magnitude lower polymerization threshold intensities than conventional ultraviolet-sensitive initiators. 23