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Label-free bacterial imaging with deep-UV-laser-induced native fluorescence.


ABSTRACT: We introduce a near-real-time optical imaging method that works via the detection of the intrinsic fluorescence of life forms upon excitation by deep-UV (DUV) illumination. A DUV (<250-nm) source enables the detection of microbes in their native state on natural materials, avoiding background autofluorescence and without the need for fluorescent dyes or tags. We demonstrate that DUV-laser-induced native fluorescence can detect bacteria on opaque surfaces at spatial scales ranging from tens of centimeters to micrometers and from communities to single cells. Given exposure times of 100 ?s and low excitation intensities, this technique enables rapid imaging of bacterial communities and cells without irreversible sample alteration or destruction. We also demonstrate the first noninvasive detection of bacteria on in situ-incubated environmental experimental samples from the deep ocean (Lo'ihi Seamount), showing the use of DUV native fluorescence for in situ detection in the deep biosphere and other nutrient-limited environments.

SUBMITTER: Bhartia R 

PROVIDER: S-EPMC2976275 | biostudies-literature | 2010 Nov

REPOSITORIES: biostudies-literature

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Label-free bacterial imaging with deep-UV-laser-induced native fluorescence.

Bhartia Rohit R   Salas Everett C EC   Hug William F WF   Reid Ray D RD   Lane Arthur L AL   Edwards Katrina J KJ   Nealson Kenneth H KH  

Applied and environmental microbiology 20100903 21


We introduce a near-real-time optical imaging method that works via the detection of the intrinsic fluorescence of life forms upon excitation by deep-UV (DUV) illumination. A DUV (<250-nm) source enables the detection of microbes in their native state on natural materials, avoiding background autofluorescence and without the need for fluorescent dyes or tags. We demonstrate that DUV-laser-induced native fluorescence can detect bacteria on opaque surfaces at spatial scales ranging from tens of ce  ...[more]

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