Journal of Zhejiang University

Journal of Zhejiang University SCIENCE A

Accepted manuscript available online (unedited version)

Two-photon polymerization based microfluidic biochip incorporating a herringbone microchannel and deterministic lateral displacement design for efficient capture of circulating tumor cells

Author(s): Xinyi LIANG, Changwei QIN, Kaiqiang LI, An REN, Jiarui HU, Weikang LV, Lunan KE, Zhen WANG, Mengfei YU, Xiuxiu JIANG, Huayong YANG, Xiaobin XU, Liang MA
Affiliation(s): State Key Laboratory of Fluid Power and Mechatronic Systems, Zhejiang University, Hangzhou 310058, China; more
Corresponding email(s): liangma@zju.edu.cn, xiaobinxu@tongji.edu.cn
Key Words: Circulating tumor cell (CTC); Cell sorting; Microfluidic chip; Two-photon polymerization; Deterministic lateral displacement

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Xinyi LIANG, Changwei QIN, Kaiqiang LI, An REN, Jiarui HU, Weikang LV, Lunan KE, Zhen WANG, Mengfei YU, Xiuxiu JIANG, Huayong YANG, Xiaobin XU, Liang MA. Two-photon polymerization based microfluidic biochip incorporating a herringbone microchannel and deterministic lateral displacement design for efficient capture of circulating tumor cells[J]. Journal of Zhejiang University Science A,in press.Frontiers of Information Technology & Electronic Engineering,in press.https://doi.org/10.1631/jzus.A2500379

@article{title="Two-photon polymerization based microfluidic biochip incorporating a herringbone microchannel and deterministic lateral displacement design for efficient capture of circulating tumor cells",
author="Xinyi LIANG, Changwei QIN, Kaiqiang LI, An REN, Jiarui HU, Weikang LV, Lunan KE, Zhen WANG, Mengfei YU, Xiuxiu JIANG, Huayong YANG, Xiaobin XU, Liang MA",
journal="Journal of Zhejiang University Science A",
year="in press",
publisher="Zhejiang University Press & Springer",
doi="https://doi.org/10.1631/jzus.A2500379"
}

%0 Journal Article
%T Two-photon polymerization based microfluidic biochip incorporating a herringbone microchannel and deterministic lateral displacement design for efficient capture of circulating tumor cells
%A Xinyi LIANG
%A Changwei QIN
%A Kaiqiang LI
%A An REN
%A Jiarui HU
%A Weikang LV
%A Lunan KE
%A Zhen WANG
%A Mengfei YU
%A Xiuxiu JIANG
%A Huayong YANG
%A Xiaobin XU
%A Liang MA
%J Journal of Zhejiang University SCIENCE A
%P 334-347
%@ 1673-565X
%D in press
%I Zhejiang University Press & Springer
doi="https://doi.org/10.1631/jzus.A2500379"

TY - JOUR
T1 - Two-photon polymerization based microfluidic biochip incorporating a herringbone microchannel and deterministic lateral displacement design for efficient capture of circulating tumor cells
A1 - Xinyi LIANG
A1 - Changwei QIN
A1 - Kaiqiang LI
A1 - An REN
A1 - Jiarui HU
A1 - Weikang LV
A1 - Lunan KE
A1 - Zhen WANG
A1 - Mengfei YU
A1 - Xiuxiu JIANG
A1 - Huayong YANG
A1 - Xiaobin XU
A1 - Liang MA
J0 - Journal of Zhejiang University Science A
SP - 334
EP - 347
%@ 1673-565X
Y1 - in press
PB - Zhejiang University Press & Springer
ER -
doi="https://doi.org/10.1631/jzus.A2500379"

Abstract
Chinese Summary
Academic Network
Reviewer Comment

Abstract: Circulating tumor cells (CTCs) are cells that become detached from a primary tumor and enter the vascular or lymphatic system. These cells contain nearly the entire genetic information of the primary tumor. Enrichment and detection of CTCs play a crucial role in prognostications and risk assessments of tumor metastasis and recurrence, evaluation of efficacy and potential medications for precision tumor therapy, and detection of dynamic biomarkers during tumor treatment. Current methods of CTC capture often face the challenge of balancing capture rate and purity. To address these issues, we propose a microfluidic biochip based on the principle of immunoaffinity, which incorporates a herringbone microchannel and deterministic lateral displacement (DLD) technology for the capture of CTCs. By manipulating the internal structural design of the microfluidic chip, we optimized the flow field within the chip, thereby enhancing the contact frequency between cells and aptamers and ultimately improving the capture rate. The proposed chip demonstrated a capture efficiency of approximately 91.87% for human breast cancer cells (MCF7), with a release rate of 77.5%. The relative activity of the released cells was approximately 94.08%. Notably, the specificity of the aptamers toward tumor cell surface antigens enables high-purity capture. Additionally, the use of DNA enzymes to digest aptamers facilitates the release of high-activity CTCs, offering a method to simultaneously achieve a high capture rate, purity, and activity enrichment.

一种基于双光子聚合的结合人字形微通道和确定性横向位移原理的微流控芯片用于循环肿瘤细胞的高效捕获

作者：梁心怡^1,2，秦昌玮^1,2，李凯强³，任安^1,2，胡家睿^1,2，吕为康^1,2，柯路南^1,2，王震³，俞梦飞⁴，江秀秀⁵，杨华勇^1,2，许晓斌⁶，马梁^1,2
机构：¹浙江大学，流体动力基础件与机电系统全国重点实验室，中国杭州，310058；²浙江大学，机械工程学院，中国杭州，310058；³浙江省人民医院输血内科变态反应中心检验医学中心，杭州医学院附属人民医院，中国杭州，310014；⁴浙江大学医学院附属口腔医院，中国杭州，310003；⁵浙江省妇科临床研究中心，浙江省母婴健康重点实验室，浙江大学医学院附属妇女医院计划生育科，中国杭州，310006；⁶同济大学，材料科学与工程学院，中国上海，201804
目的：循环肿瘤细胞（CTC）的富集和检测对肿瘤转移复发评估、疗效监测和用药指导至关重要，但现有方法难以兼顾高捕获率和高纯度。本文旨在开发一种微流控芯片，通过结合人字形微流道和确定性横向位移技术，实现循环肿瘤细胞的高捕获率、高纯度和高活性富集与释放。
创新点：1.结合人字形微流道和确定性横向位移技术的微流控芯片设计，基于免疫结合原理，并通过结构优化提高细胞与适体的接触频率，从而提升捕获率；2.利用适体对肿瘤细胞表面抗原的特异性实现高纯度捕获，并通过DNA酶消化适体实现细胞的高活性释放；3.采用计算流体力学仿真和双光子聚合打印技术，实现对芯片结构的精确设计与制作。
方法：1.通过计算流体力学仿真，优化芯片顶部人字形微流道的间距、槽宽和单元宽度，以及底部确定性横向位移柱列的柱形设计，以提高流场效率和细胞碰撞频率（图2和4）；2.采用软光刻技术加工顶部结构，探索双光子聚合打印工艺制作底部结构，并通过键合组装和适体修饰，确定最佳修饰浓度和流速承载能力（图3和5）；3.通过实验表征芯片性能：确定最佳捕获流速，验证结构优化有效性，测试释放后细胞活性和增殖能力，并评估阴性细胞捕获率和混合细胞中阳性细胞捕获纯度（图6）。
结论：1.芯片实现了对循环肿瘤细胞的高效捕获（MCF7细胞捕获率约91.87%）、高活性释放（释放率约77.57%，细胞相对活性约94.08%）以及高纯度富集（对K562阴性细胞捕获率约4.83%，混合细胞中阳性细胞捕获纯度约96.83%）；2.结合人字形微流道和确定性横向位移技术的设计，通过仿真优化有效提高了捕获性能。

关键词组：循环肿瘤细胞；细胞分选；微流控芯片

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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