Development and characterization of hollow microprobe array as a potential tool for versatile and massively parallel manipulation of single cells
Development and characterization of hollow microprobe array as a potential tool for versatile and massively parallel manipulation of single cells
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DOI：
10.1007/s10544-015-9943-z
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发表时间：
2015-04-01
影响因子：
2.8
通讯作者：
Shibata, Takayuki
中科院分区：
文献类型：
--
作者：
Nagai, Moeto;Oohara, Kiyotaka;Shibata, Takayuki
单细胞并行操作对于重建体内细胞微环境和研究细胞功能具有重要意义。为了操纵单个细胞并重建它们的环境，开发一种通用的操纵工具是必要的。在这项研究中，我们利用微型机电系统制造技术开发了一系列空心探针，并演示了对单个细胞的操作。我们用玻璃吸管进行了细胞抽吸实验，并使用标准的线性实体模型对细胞进行建模，为设计用于微创单细胞操作的中空阶梯状探针提供了信息。我们在两面都刻蚀了一片硅片，并形成了具有阶梯结构的通孔。通过等离子体增强化学气相沉积的二氧化硅沉积来减小孔洞的内径，以将细胞捕获在针尖上。该制造工艺使得能够控制探头的壁厚、内径和外径。利用所制造的探针，以平行的方式在单细胞水平上操纵单细胞并将其放置在微孔中。我们研究了不同吸入和释放压力下细胞的捕获、释放和存活率，发现细胞捕获率与吸入压力成正比，而释放速率和存活率随着吸入压力的增加而下降。所提出的操作系统使将细胞放置在良好的阵列中并观察细胞的黏附、扩散、培养和死亡成为可能。该系统有可能成为大规模并行操作和三维异质细胞分析的工具。
Parallel manipulation of single cells is important for reconstructing in vivo cellular microenvironments and studying cell functions. To manipulate single cells and reconstruct their environments, development of a versatile manipulation tool is necessary. In this study, we developed an array of hollow probes using microelectromechanical systems fabrication technology and demonstrated the manipulation of single cells. We conducted a cell aspiration experiment with a glass pipette and modeled a cell using a standard linear solid model, which provided information for designing hollow stepped probes for minimally invasive single-cell manipulation. We etched a silicon wafer on both sides and formed through holes with stepped structures. The inner diameters of the holes were reduced by SiO2 deposition of plasma-enhanced chemical vapor deposition to trap cells on the tips. This fabrication process makes it possible to control the wall thickness, inner diameter, and outer diameter of the probes. With the fabricated probes, single cells were manipulated and placed in microwells at a single-cell level in a parallel manner. We studied the capture, release, and survival rates of cells at different suction and release pressures and found that the cell trapping rate was directly proportional to the suction pressure, whereas the release rate and viability decreased with increasing the suction pressure. The proposed manipulation system makes it possible to place cells in a well array and observe the adherence, spreading, culture, and death of the cells. This system has potential as a tool for massively parallel manipulation and for three-dimensional hetero cellular assays.