The Use of Fourier Transform (FT) Surface-Enhanced Raman Scattering for Biochemical Analysis

Abstract

Surface-enhanced Raman scattering (SERS) is a powerful spectroscopic technique that has created exciting opportunities in the field of bioanalytical chemistry, where it combines ultrasensitive detection of biologically relevant molecules with vibrational spectroscopy. Due to the difficulties in preparing reproducible SERS active substrates, SERS has been mainly used as a qualitative tool. In order for SERS to be utilized as a viable tool for quantitative analysis, simple, facile SERS substrates which generate clean, highly reproducible signals must be developed. This dissertation deals with the evaluation of three different methods of SERS that led to the development of a novel substrate for the analysis of biological molecules. In this dissertation, I demonstrated the generation of a SERS signal from pyridine adsorbed to the surface of a roughened silver electrode. The Raman signal was detected utilizing a remote fiber optic probe dispersive Raman instrument. The necessary parameters for generating the optimal SERS signal were established. The SERS spectrum of pyridine was successfully characterized. The synthesis of silver colloid solutions for use as a SERS active substrate was demonstrated. Two different synthetic methods were utilized and the conditions for optimal SERS signal strength were established. SERS signals from solutions of tryptophan were successfully generated and detected using an FT-Raman spectrometer. In this dissertation. I report the successful chemical deposition of a thin silver film on the surface of a quartz/glass fiber filter for use as a SERS active substrate. The successful detection of the SERS signal of solutions of phenylalanine and tryptophan utilizing the silver-coated filters and a FT-Raman instrument was demonstrated. Significant progress was made to develop a quantifiable SERS method utilizing the silver-coated filters. The silver-coated filter SERS method presented here is a novel and promising method for biochemical analysis. The demonstrated ability to detect amino acids and mixtures of amino acids can be further extended to the detection of similar biological molecules such as proteins, tumor markers, and nucleic acids. The use of a FT-Raman instrument combined with this new SERS method allows for a method with both high sensitivity and selectivity.