UV-VIS spectroscopy is a powerfull qualitative and quantitative technique used in analytical chemistry, which gives information about electronic transitions of electrons in molecular orbitals. As in UV-VIS spectra there is no direct information on characteristic organic groups, vibrational spectroscopy (e.g. infrared) has been preferred for biological applications. In this research, we try to use state-of-the-art fiber optics probes to obtain UV-VIS-SWNIR diffusive reflectance measurements of yeasts and bacteria colonies on plate count agar in the region of 200-1200nm; in order to discriminate the following microorganisms: i) yeasts: Saccharomyces cerevisiae, Saccharomyces bayanus, Candida albicans, Yarrowia lipolytica; and ii) bacteria: Micrococcus luteus, Pseudomonas fluorescens, Escherichia coli, Bacillus cereus. Spectroscopy results show that UV-VIS-SWNIR has great potential for identifying microorganisms on plate count agar. Scattering artifacts of both colonies and plate count agar can be significantly removed using a robust mean scattering algorithm, allowing also better discriminations between the scores obtained by singular value decomposition. Hierarchical clustering analysis of UV-VIS and VIS-SWNIR decomposed spectral scores lead to the conclusion that the use of VIS-SWNIR light source produces higher discrimination ratios for all the studied microorganisms, presenting great potential for developing biotechnology applications.
Abstract:In this manuscript we explore the feasibility of using LWUV-VIS-SWNIR (340 -1100 nm) spectroscopy to classify Saccharomyces cerevisiae colony structures in YP agar and YPD agar, under different growth conditions, such as:Results show that LWUV-VIS-SWNIR spectroscopy has the potential for yeasts metabolic state identification once the spectral signatures of colonies differs from each others, being possible to acheive 100% of classification in UV-VIS and VIS-SWNIR. The UV-VIS region present high discriminant information (350-450 nm), and different responses to UV excitation were obtained. Therefore, high precision is obtained because UV-VIS and VIS-NIR exhibit different kinds of information. In the future, high precision analytical chemistry techniques such as mass spectroscopy and molecular biology transcriptomic studies should be performed in order to understand the detailed cell metabolism and genomic phenomena that characterize the yeast colony state.
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