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A Microfluidic Platform for Profiling Biomechanical Properties of Bacteria

机译:用于分析细菌生物力学特性的微流体平台

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摘要

The ability to resist mechanical forces is necessary for the survival and division of bacteria and has traditionally been probed using specialized, low-throughput techniques such as atomic force microscopy and optical tweezers. Here we demonstrate a microfluidic technique to profile the stiffness of individual bacteria and populations of bacteria. The approach is similar to micropipette aspiration used to characterize the biomechanical performance of eukaryotic cells. However, the small size and greater stiffness of bacteria relative to eukaryotic cells prevents the use of micropipettes. Here we present devices with sub-micron features capable of applying loads to bacteria in a controlled fashion. Inside the device, individual bacteria are flowed and trapped in tapered channels. Less stiff bacteria undergo greater deformation and therefore travel further into the tapered channel. Hence, the distance traversed by bacteria into a tapered channel is inversely related to cell stiffness. We demonstrate the ability of the device to characterize hundreds of bacteria at a time, measuring stiffness at 12 different applied loads at a time. The device is shown to differentiate between two bacterial species, E. coli (less stiff) and B. subtilis (more stiff), and detect differences between E. coli submitted to antibiotic treatment from untreated cells of the same species/strain. The microfluidic device is advantageous in that it requires only minimal sample preparation, no permanent cell immobilization, no staining/labeling and maintains cell viability. Our device adds detection of biomechanical phenotypes of bacteria to the list of other bacterial phenotypes currently detectable using microchip-based methods and suggests the feasibility of separating/selecting bacteria based on differences in cell stiffness.
机译:抵抗机械力的能力是细菌生存和分裂所必需的,并且传统上已使用专门的低通量技术(例如原子力显微镜和光学镊子)进行探测。在这里,我们演示了一种微流控技术,可以对单个细菌和细菌种群的硬度进行分析。该方法类似于用于表征真核细胞生物力学性能的微量移液器抽吸。但是,相对于真核细胞,细菌的体积小且硬度高,因此无法使用微量移液器。在这里,我们介绍了具有亚微米级功能的设备,这些设备能够以受控方式向细菌施加负载。在设备内部,单个细菌会流动并截留在锥形通道中。刚度较小的细菌会发生较大的变形,因此会进一步进入锥形通道。因此,细菌穿越到锥形通道的距离与细胞刚度成反比。我们展示了该设备一次可表征数百种细菌的能力,可一次测量12种不同施加负载下的硬度。该设备显示出可以区分两种细菌种类:大肠杆菌(较硬)和枯草芽孢杆菌(较硬),并检测未经抗生素处理的相同物种/菌株的大肠杆菌之间的差异。微流体装置的优点在于,其仅需要最少的样品制备,不需要永久的细胞固定,不需要染色/标记并且维持细胞活力。我们的设备将细菌生物力学表型的检测添加到了目前使用基于微芯片的方法可检测到的其他细菌表型的列表中,并提出了基于细胞硬度差异来分离/选择细菌的可行性。

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