The proposed research will use multiparameter fluorescence techniques to achieve rapid, specific detection and identification of bacteria from laboratory or clinical isolates. Several components of the proposed research will involve the use of a video fluorometer (VF), a highly efficient instrument which simultaneously records both excitation and emission spectra with a multichannel detector. The VF can acquire up to 200 emission spectra excited at up to 200 different wavelengths in less than one second. Several fluorescent species can be detected simultaneously taking advantage of the inherrently multicomponent nature of fluorescence. In this proposed study, the VF will be used to explore identification procedures with fluorescent tagged antibodies specific for a given bacteria and with identification procedures involving a series of bacterial specific enzymes and fluorescent tagged substrates. In addition, the VF will be used to continue studies involving multiple dye fingerprinting procedures where the adsorption characteristics of a bacterium for a mixture of dyes is measured and used to categorize the bacterium. The development of data processing techniques will continue. For example, two-dimensional Fourier Transform based pattern recognition techniques will be used in conjunction with Mixed-Dye-Fluorometry. Other studies will explore the use of fluorescence detected circular dichroism (FDCD) and fluorescence lifetime to provide expanded information about fluorescent components of many bacterial types.
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