基于侧向磁泳的微流控芯片对两种磁性纳米球的同时分选

doi:10.7503/cjcu20200080

摘要/Abstract

摘要：

基于磁性纳米球在微流控芯片上的侧向磁泳, 利用微流控芯片分选了不同磁响应性的磁球. 提出了包含磁性纳米球聚集与偏移的理论模型, 用于分析磁球在芯片上的侧向位移. 在理论分析的基础上设计了芯片系统, 使不同磁响应性的磁纳米球可以在芯片系统上依次被分选. 实验结果表明, 2种磁性纳米球的分选效率均可接受, 且实验操作简单; 磁响应性强的磁球可被完全分离, 这对于珍贵分析样品的分选很有价值. 该分选系统被成功用于同时分选样品中乙型肝炎病毒的DNA与丙型肝炎病毒的反转录DNA, 在生化分析中具有广阔的应用前景.

关键词: 微流控芯片, 磁性纳米球, 磁分选

Abstract:

Magnetic separation is widely used in biochemical analysis. The separations of multiple small targets by magnetic force still remain a challenge. To solve the problem, different types of magnetic probes coupled with small targets have to be sorted. Here, we propose a strategy to separate magnetic nano-beads(MNs) with either a strong or weak magnetic responsiveness on a chip system. The sorting principle is utilizing the lateral magnetophoresis of MNs. A theoretical model is proposed to analyze the lateral displacement of MNs including aggregation and deflection. On this basis, chips were designed to separate MNs. By adjusting the flow rates, MNs with different magnetic responsiveness were sequentially separated. The sorting efficiency is acceptable and the procedure is easy to perform. MNs with strong magnetic responsiveness can be completely separated, which is valuable for rare targets sorting. The system was applied to sort DNA from hepatitis B virus(HBV) and complementary DNA from hepatitis C virus(HCV) simultaneously. This system has the potential for wide application in biochemical analysis.

Key words: Microfluidic chip, Magnetic nanospheres, Magnetic separation

中图分类号:

O652.6

TrendMD:

张作然, 张利, 张志凌. 基于侧向磁泳的微流控芯片对两种磁性纳米球的同时分选. 高等学校化学学报, 2020, 41(6): 1243.

ZHANG Zuoran, ZHANG Li, ZHANG Zhiling. On-chip Sorting of Beads with Different Magnetic Responsiveness by Lateral Magnetophoresis ^†. Chem. J. Chinese Universities, 2020, 41(6): 1243.

图/表 9

Table 1 Sequences of DNA

DNA	Sequence
Capture-HBV	Biotin-TEG-5'-TGGCTTTCAGTTATA-3'
Capture-HCV	Biotin-TEG-5'-TCGTGGATAAACCCG-3'
Target-HBV	5'-ATACCACATCATCCATATAACTGAAAGCCA-3'
Target-HCV	5'-ATCTCCAGGCATTGAGCGGGTTTATCCACGA-3'
Probe-HBV	5'-TGGATGATGTGGTAT-3'-TEG-biotin
Probe-HCV	5'-CTCAATGCCTGGAGAT-3'-TEG-biotin

Scheme 1 Schematic of microfluidic chip system

Fig.1 Characterization of beads with strong magnetic responsiveness(5-MNs) and weak magnetic responsiveness(1-MNs) (A, D) TEM images; (B, E) particle size statistics of 1-MNs(A, B) and 5-MNS(D, E); (C) magnetic hysteresis loops for 5-MNs(a) and 1-MNs(b); (F) fluorescence spectra of 1-GMNs(a) and 5-RMNs(b).

Fig.2 Simulation result of the magnetic field gradient(?B) around Ni strips Each Ni strips was 50 μm in width and 11 μm in height. The interval between Ni strips was 100 μm. The thickness of ITO glass under the Ni strips was 1.1 mm. Ni strips were covered by an extremely thin PDMS layer. The fluid channel was above the Ni strips.

Fig.3 Motion analysis of MNs (A) Velocity analysis of a single MNs; (B) the motion of magnetic composited particles.

Fig.4 On-chip movement of 5-RMNs and 1-GMN at different flow rates (A) Structure of the chip; (B)—(F) images were taken in a bright field; (B) without magnetic field, MNs did not move along the Ni strips; (C) when an external magnetic field was added, the MNs became magnetized and clumped into visible micro-sized particles; (D)—(F) images of the motion of MNs at different flow rates; (G)—(I) fluorescence field images and fluorescence spectra of the fluorescent MNs collected at each outlet.

Fig.5 Fluorescence spectra of samples The flow rate of the sample was 5 μL/min and of the buffer was 30 μL/min. (A) Sample with only 1-RMNs; (B)—(D) samples with 5-MNs/1-RMNs mass ratios of 1:1, 2:1, 1:2, respectively. a. Outlet 1; b. outlet 2.

Fig.6 Separation efficiency of 5-RMNs and 1-GMNs Outlet 1 collected 5-MNs and outlet 2 collected 1-MNs. (A), (B) Separation efficiency of 5-RMNs and 1-GMNs by chip 1. (A) Flow rates: sample 5 μL/min, buffer 30 μL/min; (B) flow rates: sample 5 μL/min, buffer 32 μL/min; (C) relative separation percentages of 1-GMNs and 5-RMNs with flow rates by chip 1. Standard deviation was calculated from three experimental runs; (D), (E) separation efficiency of 1-GMNs by chip 2; (D) flow rates: sample 5 μL/min, buffer 5 μL/min; (E) flow rates: sample 5 μL/min, buffer 10 μL/min; (F) separation efficiency of 5-RMNs and 1-GMNs for the chip system. a. Outlet 1; b. outlet 2; c. waste.

Fig.7 Separation of target DNA of HBV and HCV from the sample Distributions of HBV DNA(A) and HCV complementary DNA(B) at outlet 1(a), outlet 2(b) and the waste outlet(c).

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