Raman Microspectroscopy for Species Identification andMapping within Bacterial Biofilms 英文参考文献.docVIP

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Raman Microspectroscopy for Species Identification andMapping within Bacterial Biofilms 英文参考文献.doc

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Raman Microspectroscopy for Species Identification andMapping within Bacterial Biofilms 英文参考文献

Beieretal.AMBExpress2012,2:35 /content/2/1/35 ORIGINAL ARTICLE OpenAccess Ramanmicrospectroscopyforspecies identi?cationandmappingwithinbacterial bio?lms BrookeDBeier1,RobertGQuivey2andAndrewJBerger1* Abstract Anewmethodofmappingmultiplespeciesoforalbacteriainintactbio?lmshasbeendeveloped,usingtheoptical techniqueofconfocalRamanmicroscopy.Aspeciesclassi?cationalgorithm,developedondriedbio?lms,wasusedto analyzespectraofhydratedbio?lmscontainingtwomicrobialspeciescentraltodentalhealth:Streptococcussanguinis andStreptococcusmutans.Thealgorithmtransferredsuccessfullytothehydratedenvironment,correctlyidentifying thespeciesoforiginofsingle-speciesbio?lms.Wethenusedthealgorithmsuccessfullybothtodetectthepresence oftwospeciesinmixedbio?lmsandtocreatespatialmapswithinthesebio?lms. Keywords: Ramanspectroscopy,Confocalmicroscopy,Bio?lms,Bacteria,Dentalplaque Introduction inintact,un?xedspecimensinalabel-free,non-contact The identi?cation of microbial species within sample manner would therefore be valuable. Such a technique specimensisrelevanttoboththemicrobiologicalresearch couldalsomakelongitudinalstudyofsampleslikebio?lms laboratory and the clinical setting. A number of stan- apossibility. dardmethodsofspeciesidenti?cationarecurrentlyused. Optical spectroscopy o?ers the ability to acquire Selectiveplatingmaybeusedtoidentifytheconstituent chemically-speci?c information at micron-scale resolu- speciesandtoobtainanorder-of-magnitudeestimateof tionwithoutsamplecontact.OnesuchmethodisRaman the initial concentrations of bacteria present in a sam- spectroscopy,whichhasbeenusedextensivelyforbiolog- ple. Since this method involves serial dilutions, it can ical applications. Raman spectroscopy detects the pres- takeuptoseveraldaysandisinsensitivetocellsthatare ence of molecular bonds via the inelastic scattering of deadorincapableofreproductionbythetimethesam- laserlight;eachmoleculeimpartsadi?erent“?ngerprint pleisprocured. Quantitativepolymerasec