Targeted cell immobilization by ultrasound microbeam.
Targeted cell immobilization by ultrasound microbeam.
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DOI：
10.1002/bit.23073
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发表时间：
2011-07
影响因子：
3.8
通讯作者：
Shung, K. Kirk
中科院分区：
文献类型：
--
作者：
Lee, Jungwoo;Lee, Changyang;Kim, Hyung Ham;Jakob, Anette;Lemor, Robert;Teh, Shia-Yen;Lee, Abraham;Shung, K. Kirk
已经开发了对细胞施加机械应力的各种技术来研究细胞对外部控制刺激的反应。基本的机械力传导过程，即施加的物理力如何转化为生化信号，通常通过将这些力通过细胞传递并在细胞水平上探测其途径来研究。事实上，许多细胞生物力学研究已经通过捕获（或固定）单个细胞来进行，无论是附着在固体基质上还是悬浮在液体介质中。在这种情况下，我们展示了二维声捕获，其中直径为125 μm的脂滴横向指向焦点（或陷阱中心），类似于光镊。在30 MHz聚焦超声波束产生的恢复力的影响下，捕获的液滴表现得好像被线性弹簧拴在焦点上。为了将这种方法应用于米氏体系（细胞直径>波长）中的细胞操纵，具有接近细胞尺寸的波束宽度的声束的可用性是至关重要的。这只能在高于100 MHz的频率下实现。我们将频率范围从100 MHz到几GHz的超声波束定义为超声波微束，因为焦点处的横向波束宽度将在微米范围内（评审员#1）。因此，我们设计并制作了一个能发射200 MHz聚焦声束的氧化锌（ZnO）换能器，并将一个10 μm的人白血病细胞（K-562）置于陷阱中。通过在焦平面上机械平移换能器，使细胞相对于陷阱中心横向移位。在二维空间中探测被捕获细胞的横向位移和位置轨迹，表明这些细胞的缩回运动与30 MHz下的脂滴相似。讨论了该工具用于研究白色血细胞和内皮细胞之间的细胞粘附的潜力，表明其作为单细胞操纵器的能力。
Various techniques exerting mechanical stress on cells have been developed to investigate cellular responses to externally controlled stimuli. Fundamental mechanotransduction processes, how applied physical forces are converted into biochemical signals, have often been examined by transmitting such forces through cells and probing its pathway at cellular levels. In fact, many cellular biomechanics studies have been performed by trapping (or immobilizing) individual cells, either attached to solid substrates or suspended in liquid media. In that context, we demonstrated two-dimensional acoustic trapping, where a lipid droplet of 125 μm in diameter was directed transversely towards the focus (or the trap center) similar to that of optical tweezers. Under the influence of restoring forces created by a 30 MHz focused ultrasound beam, the trapped droplet behaved as if tethered to the focus by a linear spring. In order to apply this method to cellular manipulation in the Mie regime (cell diameter > wavelength), the availability of sound beams with its beamwidth approaching cell size is crucial. This can only be achieved at a frequency higher than 100 MHz. We define ultrasound beams in the frequency range from 100 MHz to a few GHz as ultrasound microbeams because the lateral beamwidth at the focus would be in the micron range (reviewer #1). Hence a zinc oxide (ZnO) transducer that was designed and fabricated to transmit a 200 MHz focused sound beam was employed to immobilize a 10 μm human leukemia cell (K-562) within the trap. The cell was laterally displaced with respect to the trap center by mechanically translating the transducer over the focal plane. Both lateral displacement and position trajectory of the trapped cell were probed in a two-dimensional space, indicating that the retracting motion of these cells was similar to that of the lipid droplets at 30 MHz. The potential of this tool for studying cellular adhesion between white blood cells and endothelial cells was discussed, suggesting its capability as a single cell manipulator.
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