| This doctoral project is focused on the implementation of a novel micron and sub-micron particle characterization technology for in-situ, continuous monitoring and detecting of microorganisms in water. The particle technology is based on simultaneous characterizing the joint particle property distribution (size, shape, and chemical composition) through the combined fractionation/separation and light scattering detection and interpretation techniques.; Over more than a decade, field-flow fractionation (FFF) has shown to be well-suited for the separation and/or selection of bacteria (Giddings, 1993). As the most universal fractionation technique among the FFF family, flow field-flow fractionation (FFFF) has been chosen as the separation device in this research. The multi-angle laser light scattering (MALLS) photometer and the UV-vis/liquid core optical waveguide constitute the primary on-line light scattering detection system. The angular spectra obtained by the MALLS photometer provided information on the shape of microorganism; the multi-wavelength transmission spectra of microorganisms contain quantitative information on their size, number, shape, chemical composition and internal structure, which are essential for identification and classification of microorganisms.; Both experimental results and the theoretical prediction have revealed that the particle size resolution capabilities of the FFFF fractionation system coupled with the sensitivity of the laser light scattering to particle shape, and the sensitivity of the UV-vis spectra to cell size, shape, cell orientation and chemical composition offer an integrated system for the identification and classification of microorganisms. The ability to discriminate between cell species was demonstrated by the light scattering and absorption interpretation model, which is based on light scattering theory (Rayleigh-Debye-Gans approximation), spectral deconvolution techniques, and on the approximation of the frequency dependent optical properties of the basic constituents of microorganisms. It is further demonstrated that the combined multi-angle light scattering and multi-wavelength transmission spectroscopic technique can provide reliable information on the cell size, cell count, shape, cell alignment, chemical composition and basic understanding of the light scattering origins within the cell. Advances in the development of miniaturized spectrometers and the micromachined field-flow fractionation technique increase the potential of this method as a candidate for a rapid, reliable and efficient biosensor. |
