A High-Throughput Automated Microinjection System for Human Cells With Small Size
A High-Throughput Automated Microinjection System for Human Cells With Small Size
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DOI：
10.1109/tmech.2015.2476362
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发表时间：
2016-04-01
影响因子：
6.4
通讯作者：
Sun, Dong
中科院分区：
文献类型：
--
作者：
Chow, Yu Ting;Chen, Shuxun;Sun, Dong
本文介绍了一种高生产率的小细胞自动显微注射系统的开发。与许多现有的显微注射系统，主要是设计用于相对大规模的生物样品相比，所报道的技术将能够自动注射到直径小于25 μ m的细胞，这是许多人类细胞的典型尺寸。微流控技术已被用于设计一种基于真空的细胞固定装置，用于固定细胞。细胞保持器采用数百个规则形状的通道，并且每个通道被一个细胞占据。采用基于视觉的位置跟踪方法对靶细胞进行自动识别和定位，利用边缘模板匹配法测量细胞位置，并利用明场图像简化识别过程。利用一个装有微量吸管的三自由度微机器人系统进行细胞注射，注射速度为35个细胞/min，注射精度为0.2 μ m。对人包皮成纤维细胞和人胚胎干细胞衍生的心室心肌细胞进行注射实验以证明所开发的系统的有效性。超过1000个细胞被注射了荧光标记物。实验结果表明，开发的注射系统具有较高的生产率和准确性，并在注射后的细胞存活率良好。
This paper presents the development of an automated microinjection system with high productivity for small cells. Compared with many existing microinjection systems that are primarily designed for relatively large-scaled biological samples, the reported technology will enable an automated injection into the cells with a diameter smaller than 25 mu m, which is the typical size of many human cells. Microfluidic technology has been used to design a vacuum-based cell-holding device for immobilizing cells. The cell holder employs hundreds of regular-shaped channels, and each channel is occupied by one cell. The vision-based position-tracking method is used to recognize and position the target cells automatically, in which the cell position is measured by edge template-matching method, and a bright field image is used to simplify the recognition process. A 3-DOF microrobotic system mounted with a micropipette is used to conduct cell injection task with the speed of 35 cells/min and the precision of 0.2 mu m. Injection experiments on human foreskin fibroblast and human embryonic stem cell-derived ventricular cardiomyocyte were performed to demonstrate the effectiveness of the developed system. More than 1000 cells were injected with fluorescent markers. Experimental results showed that the developed injection system exhibits high productivity and accuracy, and results in a good cell survival rate after injection.