基于热发泡技术的高通量单细胞打印  

High Throughput Single-Cell Printing Based on Thermal Bubble Technology

作　　者：周钦 王琨 关一民 黄鹏 朱莉莎 王秋实 金文最 刘德盟 Zhou Qin;Wang Kun;Guan Yimin;Huang Peng;Zhu Lisha;Wang Qiushi;Jin Wenzui;Liu Demeng(School of Microelectronics,Shanghai University,Shanghai 201800,China;Shanghai Aure Technology Limited Company,Shanghai 201800,China)

机构地区：[1]上海大学微电子学院,上海201800 [2]上海傲睿科技有限公司,上海201800

出　　处：《微纳电子技术》2023年第10期1669-1676,共8页Micronanoelectronic Technology

基　　金：国家自然科学基金(82130069);上海市重大市级科技项目(2021SHZDZX)。

摘　　要：传统单细胞分离技术包含流式细胞术、激光捕获法和显微操作法等。然而,流式细胞术需要大量样本,且仪器体积庞大,价格昂贵;激光捕获法和显微操作法则耗时长,并且单细胞获取效率和通量较低。将热发泡喷墨技术应用于单细胞打印。利用热发泡喷嘴驱动细胞悬液,因此无需外接注射泵,并在芯片上集成大量喷嘴,实现高通量单细胞打印。首先通过Open CV算法分割喷嘴图像,然后通过卷积神经网络(CNN)对带有单细胞的喷嘴图像进行训练和识别,最后控制喷嘴实现单细胞打印。实现了对浓度1×10^(6)cells/m L的CHO-K1+CDCHO和CHO-K1+DMEM/F12+FBS两种细胞悬液的16块96孔板单细胞打印,单个96孔板分选在5 min内完成。两种细胞悬液即时细胞活性损耗分别为9.1%和8.3%,单细胞打印率分别达到86.7%和87.3%,单细胞克隆率分别达到44.5%和36.9%。Traditional single-cell isolation techniques include flow cytometry,laser capture method and micromanipulation method,etc.Flow cytometry requires a large number of samples,and the instrument is large and expensive,while laser capture and micromanipulation methods are time-consuming and have low single-cell acquisition efficiency and throughput.The thermal bubble inkjet technology was applied to single-cell printing.The thermal bubble nozzle was utilized to drive the cell suspension,eliminating the need for an external syringe pump,and a large number of nozzles were integrated on the chip to achieve high-throughput single-cell printing.Firstly,the nozzle images was segmented by OpenCV algorithm,and then nozzle images with single-cell were trained and recognized by convolutional neural network(CNN).Finally,the single-cell printing was achieved by controlling the nozzle.The single-cell printing of 1696-well plates for CHO-K1+CDCHO and CHO-K1+DMEM/F12+FBS cell suspensions with the concentration of 1×10^(6)cells/mL was achieved.The sorting time of a single 96-well plate is less than 5 min.The real-time cell viability losses of the two cell suspensions are 9.1%and 8.3%respectively,the single-cell printing rates are 86.7%and 87.3%respectively,and the single-cell cloning rates are 44.5%and 36.9%respectively.

关 键 词：微流控 热发泡技术 单细胞打印 图像算法 单细胞克隆 

分 类 号：TN303[电子电信—物理电子学]