To guarantee safe supply of drinking water, there is a need for fast, sensitive and robust techniques for early warning of potentially pathogenic microorganisms. An optical measurement system has been developed to spectrally study the UVinduced autofluorescence from single microorganisms in a water flow. Particles in the flow are detected with a continuouswave laser whereby an ultraviolet laser pulse is fired and a spectrometer measures emitted fluorescence. Suspensions of Cryptosporidium parvum (Crypto), Bacillus atrophaeus (BG, a non-pathogenic simulant bacterial spore) and Escherichia coli (Ecoli), in distilled and tap water have been examined. The results from single particle measurements are compared to fluorescence emission spectra captured on suspensions with a fluorescence spectrometer and the strength and variability of single organism spectra, with respect to detection applications, is also investigated.
There is a need for time efficient evaluation methods to discriminate between viable and dead bacterial spores. In this work, the potential to use the autofluorescence from spore suspensions for evaluation of spore deactivation processes is investigated. Bacillus thuringiensis and Bacillus anthracis ATCC 4229 spores were exposed to UV-radiation for deactivation and the fluorescence response was monitored at different radiation doses and the deactivation was evaluated via traditional bacterial incubation on agar culture plates. For excitation wavelengths of, e.g., 280 m and 330 nm, differences in the fluorescence response could be observed for different live:dead ratios.
Spore-forming bacteria that cause diseases pose a danger in our society. When in spore form, bacteria can survive high temperatures and resist a plethora of disinfection chemicals. Effective disinfection approaches are thus critical. Since a population of bacterial spores is heterogeneous in many aspects, single spore analyzing methods are suitable when heterogeneous information cannot be neglected. We present in this work a highresolution Laser Raman optical tweezers that can trap single spores and characterize their Raman spectra. We first evaluate our system by measuring Raman spectra of spores, and purified DNA and DPA. Thereafter, we expose Bacillus thuringiensis spores to peracetic acid, chlorine dioxide, and sodium hypochlorite, which are common disinfection chemicals. The data reveals how these agents change the constitutes of a spore over time, thus improving on the mode of action of these disinfection chemicals.
Microfabricated diamond waveguides, between 5 and 20 μm thick, manufactured by chemical vapor deposition of diamond, followed by standard lithographic techniques and inductively coupled plasma etching of diamond, are used as bio-chemical sensors in the mid infrared domain: 5-11 μm. Infrared light, emitted from a broadly tunable quantum cascade laser with a wavelength resolution smaller than 20 nm, is coupled through the diamond waveguides for attenuated total reflection spectroscopy. The expected advantages of these waveguides are a high sensitivity due to the high number of internal reflections along the propagation direction, a high transmittance in the mid-IR domain, the bio-compatibility of diamond and the possibility of functionalizing the surface layer. The sensor will be used for analyzing different forms of proteins such as α-synuclein which is relevant in understanding the mechanism behind Parkinson's disease. The fabrication process of the waveguide, its characteristics and several geometries are introduced. The optical setup of the biosensor is described and our first measurements on two analytes to demonstrate the principle of the sensing method will be presented. Future use of this sensor includes the functionalization of the diamond waveguide sensor surface to be able to fish out alpha-synuclein from cerebrospinal fluid.
Results related to laser induced breakdown spectroscopy (LIBS) as an analytical tool for applications regarding CWA and BWA detection/monitoring will be presented and discussed in this paper. A ‘real-time’ aerosol analysis set-up using LIBS on single μm-sized particles (sampled from ambient air into a particle stream) has been developed and evaluated. Here, a two-stage triggering unit ensures a high hit-rate of the sampled aerosol particles and the optical emission from the laser induced plasma is collected and coupled into an echelle spectrometer equipped with an intensified CCD detector. Each CCD image (echellogram), optimally originating from a single μm-sized particle, is then analyzed and the result treated by an alarm algorithm built from a database using multivariate data analysis. The database signatures of simulant agents and interferents were obtained in controlled atmospheres (aerosol chamber/wind tunnel) as well as from measurements in different ambient background. The LIBS bioaerosol system with alarm algorithm was also tested in ‘real-life’ settings (subway station) during simulant dispersions. Painted surfaces have also been analyzed by LIBS to obtain information about residues of organophosphates on, or within, the paint. Depth analysis has been performed, which illustrated the possibility to monitor diffusion and penetration behavior of neat CWAs and simulant chemicals in the paint layer by following the intensity of phosphorous emission lines in single shot LIBS spectra as function of number of laser pulses. In addition, LIBS analysis was also performed after simple ethanol decontamination procedures, after which P emission lines still could be observed. The possibilities and challenges associated with the different set-ups and applications will be briefly discussed in connection with the presented results.
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