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Label-Free Bacterial Imaging with Deep-UV-Laser-Induced Native Fluorescence

机译:深紫外激光诱导的天然荧光的无标签细菌成像

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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 μs 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.
机译:我们介绍了一种近实时光学成像方法,该方法通过检测深紫外线(DUV)激发后生命形式的固有荧光来工作。 DUV(<250-nm)源可在天然材料上检测其天然状态的微生物,从而避免了背景自发荧光,并且无需荧光染料或标签。我们证明DUV激光诱导的天然荧光可以检测不透明表面上细菌的空间尺度,范围从几十厘米到微米,从社区到单个细胞。给定100μs的曝光时间和低激发强度,该技术可以快速成像细菌群落和细胞,而不会发生不可逆的样品改变或破坏。我们还展示了在深海(Lo'ihi Seamount)原位孵化的环境实验样品上首次无创检测细菌的方法,展示了利用DUV天然荧光在深生物圈和其他营养有限的环境中进行原位检测。

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