A microfluidic platform for profiling biomechanical properties of bacteria.
A microfluidic platform for profiling biomechanical properties of bacteria.
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DOI：
10.1039/c3lc51428e
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发表时间：
2014-07-21
期刊：
影响因子：
6.1
通讯作者：
Hernandez CJ
中科院分区：
文献类型：
--
作者：
Sun X;Weinlandt WD;Patel H;Wu M;Hernandez CJ
抵抗机械力的能力对于细菌的生存和分裂是必要的，传统上使用专门的低通量技术进行探测，如原子力显微镜和光学镊子。在这里，我们展示了一种微流体技术来描述单个细菌和细菌群体的刚度。该方法类似于用于表征真核细胞生物力学性能的微管抽吸。然而，与真核细胞相比，细菌的体积小，硬度大，这阻碍了微移液管的使用。在这里，我们提出了具有亚微米特征的设备，能够以受控的方式对细菌施加负载。在设备内部，单个细菌流动并被困在锥形通道中。硬度较低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，细菌进入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表征数百种细菌的能力，一次测量12种不同施加载荷下的刚度。该装置被证明可以区分两种细菌，大肠杆菌（较不僵硬）和枯草芽孢杆菌（较僵硬），并检测同一物种/菌株未经处理的细胞经抗生素治疗的大肠杆菌之间的差异。微流控装置的优势在于它只需要最少的样品制备，不需要永久的细胞固定，不需要染色/标记并保持细胞活力。我们的设备将细菌的生物力学表型检测添加到目前使用基于微芯片的方法可检测的其他细菌表型列表中，并表明基于细胞刚度差异分离/选择细菌的可行性。
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.
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影响因子：
10.2
作者：
Nakanishi K;Kogure A;Fujii T;Kokawa R;Deuchi K
通讯作者：
Deuchi K
影响因子：
6.1
作者：
Hulme, S. Elizabeth;Shevkoplyas, Sergey S.;Whitesides, George M.
通讯作者：
Whitesides, George M.
在杆状革兰氏阴性细菌中维持细胞形状的机制。
影响因子：
3.6
作者：
Furchtgott L;Wingreen NS;Huang KC
通讯作者：
Huang KC
影响因子：
7.4
作者：
Hawkins, Benjamin G.;Huang, Chao;Kirby, Brian J.
通讯作者：
Kirby, Brian J.
影响因子：
3.4
作者：
Herricks T;Antia M;Rathod PK
通讯作者：
Rathod PK