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海洋微生物对微尺度营养层的趋化反应。

Chemotactic response of marine micro-organisms to micro-scale nutrient layers.

作者信息

Seymour Justin R, Stocker Roman

机构信息

Environmental Microfluidics Group, Massachusetts Institute of Technology, USA.

出版信息

J Vis Exp. 2007(4):203. doi: 10.3791/203. Epub 2007 May 28.

Abstract

The degree to which planktonic microbes can exploit microscale resource patches will have considerable implications for oceanic trophodynamics and biogeochemical flux. However, to take advantage of nutrient patches in the ocean, swimming microbes must overcome the influences of physical forces including molecular diffusion and turbulent shear, which will limit the availability of patches and the ability of bacteria to locate them. Until recently, methodological limitations have precluded direct examinations of microbial behaviour within patchy habitats and realistic small-scale flow conditions. Hence, much of our current knowledge regarding microbial behaviour in the ocean has been procured from theoretical predictions. To obtain new information on microbial foraging behaviour in the ocean we have applied soft lithographic fabrication techniques to develop 2 microfluidic devices, which we have used to create (i) microscale nutrient patches with dimensions and diffusive characteristics relevant to oceanic processes and (ii) microscale vortices, with shear rates corresponding to those expected in the ocean. These microfluidic devices have permitted a first direct examination of microbial swimming and chemotactic behaviour within a heterogeneous and dynamic seascape. The combined use of epifluorescence and phase contrast microscopy allow direct examinations of the physical dimensions and diffusive characteristics of nutrient patches, while observing the population-level aggregative response, in addition to the swimming behaviour of individual microbes. These experiments have revealed that some species of phytoplankton, heterotrophic bacteria and phagotrophic protists are adept at locating and exploiting diffusing microscale resource patches within very short time frames. We have also shown that up to moderate shear rates, marine bacteria are able to fight the flow and swim through their environment at their own accord. However, beyond a threshold high shear level, bacteria are aligned in the shear flow and are less capable of swimming without disturbance from the flow. Microfluidics represents a novel and inexpensive approach for studying aquatic microbial ecology, and due to its suitability for accurately creating realistic flow fields and substrate gradients at the microscale, is ideally applicable to examinations of microbial behaviour at the smallest scales of interaction. We therefore suggest that microfluidics represents a valuable tool for obtaining a better understanding of the ecology of microorganisms in the ocean.

摘要

浮游微生物利用微观尺度资源斑块的程度对海洋营养动力学和生物地球化学通量具有重大影响。然而,为了利用海洋中的营养斑块,游动的微生物必须克服包括分子扩散和湍流剪切在内的物理力的影响,这些物理力会限制斑块的可用性以及细菌定位它们的能力。直到最近,方法上的局限性阻碍了在斑块状栖息地和现实的小尺度流动条件下对微生物行为的直接研究。因此,我们目前关于海洋中微生物行为的许多知识都是从理论预测中获得的。为了获取有关海洋中微生物觅食行为的新信息,我们应用软光刻制造技术开发了2种微流控装置,用于创建(i)具有与海洋过程相关的尺寸和扩散特征的微观尺度营养斑块,以及(ii)具有与海洋中预期剪切速率相对应的微观尺度涡旋。这些微流控装置首次允许直接研究异质动态海景中的微生物游动和趋化行为。落射荧光显微镜和相差显微镜的联合使用,除了观察单个微生物的游动行为外,还能直接检查营养斑块的物理尺寸和扩散特征,同时观察群体水平的聚集反应。这些实验表明,一些浮游植物、异养细菌和吞噬性原生生物物种能够在极短的时间内定位并利用扩散的微观尺度资源斑块。我们还表明,在中等剪切速率以下,海洋细菌能够逆流游动并自行在其环境中移动。然而,超过阈值高剪切水平后,细菌会在剪切流中排列,并且在没有水流干扰的情况下游动能力降低。微流控技术是一种研究水生微生物生态学的新颖且廉价的方法,由于其适合在微观尺度上精确创建现实的流场和底物梯度,非常适用于在最小相互作用尺度上检查微生物行为。因此,我们认为微流控技术是一个有价值的工具,有助于更好地理解海洋中微生物的生态学。

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本文引用的文献

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Microbial structuring of marine ecosystems.海洋生态系统的微生物结构
Nat Rev Microbiol. 2007 Oct;5(10):782-91. doi: 10.1038/nrmicro1747.
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Bacterial metapopulations in nanofabricated landscapes.纳米制造景观中的细菌集合种群
Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17290-5. doi: 10.1073/pnas.0607971103. Epub 2006 Nov 7.
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Influence of topology on bacterial social interaction.拓扑结构对细菌社会相互作用的影响。
Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):13910-5. doi: 10.1073/pnas.1935975100. Epub 2003 Nov 17.
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A sensitive, versatile microfluidic assay for bacterial chemotaxis.一种用于细菌趋化性的灵敏、通用的微流控检测方法。
Proc Natl Acad Sci U S A. 2003 Apr 29;100(9):5449-54. doi: 10.1073/pnas.0931258100. Epub 2003 Apr 18.
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Soft lithography in biology and biochemistry.生物学与生物化学中的软光刻技术。
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