raman microspectroscopy for species identification andmapping within bacterial biofilms细菌生物膜内拉曼显微镜物种识别和绘制.pdfVIP

Beier et al. AMB Express 2012, 2:35 /content/2/1/35 ORIGINAL ARTICLE Open Access Raman microspectroscopy for species identification and mapping within bacterial biofilms 1 2 1* Brooke D Beier , Robert G Quivey and Andrew J Berger Abstract A new method of mapping multiple species of oral bacteria in intact biofilms has been developed, using the optical technique of confocal Raman microscopy. A species classification algorithm, developed on dried biofilms, was used to analyze spectra of hydrated biofilms containing two microbial species central to dental health: Streptococcus sanguinis and Streptococcus mutans. The algorithm transferred successfully to the hydrated environment, correctly identifying the species of origin of single-species biofilms. We then used the algorithm successfully both to detect the presence of two species in mixed biofilms and to create spatial maps within these biofilms. Keywords: Raman spectroscopy, Confocal microscopy, Biofilms, Bacteria, Dental plaque Introduction in intact, unfixed specimens in a label-free, non-contact The identification of microbial species within sample manner would therefore be valuable. Such a technique specimens is relevant to both the microbiological research could also make longitudinal study of samples like biofilms laboratory and the clinical setting. A number of stan- a possibility. dard methods of species identification are currently used. Optical spectroscopy offers the ability to acquire Selective plating may be used to identify the constituent chemically-specific information at micron-scale resolu- species and to obtain an order-of-m